Anti-LAG3 antibodies and antigen-binding fragments

ABSTRACT

The present invention includes antibodies and antigen-binding fragments thereof that specifically bind to human or cynomolgous monkey LAG3 as well as immunoglobulin chains thereof and polynucleotides encoding the same along with injection devices comprising such antibodies or fragments. Vaccines including such antibodies and fragments as well as compositions comprising the antibodies and fragments (e.g., including anti-PD1 antibodies) are included in the invention. Methods for treating or preventing cancer or infection using such compositions are also provided. In addition, methods for recombinant expression of the antibodies and fragments are part of the present invention.

This Application is a divisional of U.S. application Ser. No.15/482,355, which is a continuation of U.S. application Ser. No.15/305,011, filed Oct. 18, 2016, which is a U.S. National Phaseapplication under 35 U.S.C. § 371 of PCT/US15/45481, filed Aug. 17,2015, which claims priority to U.S. Provisional Patent Application No.62/171,319, filed Jun. 5, 2015; and claims priority to U.S. ProvisionalPatent Application No. 62/039,081, filed Aug. 19, 2014; each of which isherein incorporated by reference in its entirety.

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Dec. 4, 2018, isnamed 23791USDIV-SEQTXT-4DEC2018 and is 968 kilobytes in size.

FIELD OF THE INVENTION

The present invention relates to anti-LAG3 antibodies as well as use ofthe antibodies of the present invention in the treatment of diseasessuch as cancer and infection.

BACKGROUND OF THE INVENTION

LAG3 (CD223) is a cell surface molecule expressed on activated T cells(Huard et al. Immunogenetics 39:213-217, 1994), NK cells (Triebel et al.J Exp Med 171:1393-1405, 1990), B cells (Kisielow et al. Eur J Immunol35:2081-2088, 2005), and plasmacytoid dendritic cells (Workman et al. JImmunol 182:1885-1891, 2009) that plays an important role in thefunction of these lymphocyte subsets. In addition, the interactionbetween LAG3 and its major ligand, Class II MHC, is thought to play arole in modulating dendritic cell function (Andreae et al. J Immunol168:3874-3880, 2002). Recent preclinical studies have documented a rolefor LAG-3 in CD8 T-cell exhaustion (Blackburn et al. Nat Immunol10:29-37, 2009).

As with chronic viral infection, tumor antigen-specific CD4⁺ and CD8⁺ Tcells display impaired effector function and an exhausted phenotypecharacterized by decreased production of pro-inflammatory cytokines andhyporesponsiveness to antigenic re-stimulation. This is mediated by cellextrinsic mechanisms, such as regulatory T-cells (Treg), and cellintrinsic mechanisms, such as inhibitory molecules that are upregulatedon exhausted, tumor-infiltrating lymphocytes (TIL). These inhibitorymechanisms represent a formidable barrier to effective antitumorimmunity.

LAG-is expressed on tolerized TILs suggesting that they contribute totumor-mediated immune suppression. Inhibition of LAG3 may lead toenhanced activation of antigen-specific T cells from which a therapeuticbenefit may be gained. There is a need in the art for high efficacytherapeutic antibodies which antagonize the activity of LAG3 which canbe used to generate a robust immune response to tumors.

SUMMARY OF THE INVENTION

The present invention provides an antibody or antigen-binding fragmentthereof (e.g., an antibody, an antigen-binding fragment, monoclonalantibodies, polyclonal antibodies, a multispecific antibody, a humanizedantibody, an isolated antibody or antigen-binding fragment thereof, ahumanized antagonist antibody, a fully human antibody, a chimericantibody and a camelized single domain antibody) that specifically bindsLAG3 (e.g., human and/or Macaqa fascicularis, e.g., SEQ ID NOs: 443 or445) comprising: a heavy chain immunoglobulin variable region having atleast 78.99% amino acid sequence identity to amino acids 1-119 of SEQ IDNO: 106; and/or a light chain immunoglobulin variable region having atleast 78.38% amino acid sequence identity to amino acids 1-111 of SEQ IDNO: 224.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention comprises thelight and heavy chain immunoglobulin (e.g., heavy and light chainvariable domains or heavy and light chain CDRs) of Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 or Ab9 as set forth herein.

The present invention provides an antibody or antigen-binding fragmentthereof that specifically binds LAG3 (e.g., human or cynomolgous monkeyLAG3) comprising (a) the CDR1, CDR2, and CDR3 of a V_(L) domain of animmunoglobulin chain that comprises the amino acid sequence set forth inSEQ ID NO: 7, 17, 27, 37, 57, 59, 61, 63, 65, 101, 126, 130, 132, 136,138, 208, 210, 224, 226, 228, 230, 232, 241, 257, 259, 261, 263, 351,369, 371, 373, 375, 401, 403, 405, 426, 427, 450-453 or 459-461; and/or(b) the CDR1, CDR2, and CDR3 of a V_(H) domain of an immunoglobulinchain that comprises the amino acid sequence set forth in SEQ ID NO: 2,12, 22, 32, 45, 47, 49, 51, 53, 55, 67, 69, 71, 73, 75, 77, 79, 81, 83,85, 87, 89, 91, 93, 95, 97, 99, 103, 106, 108, 110, 112, 114, 116, 118,120, 122, 124, 128, 134, 140, 142, 144, 146, 148, 150, 152, 154, 156,158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184,186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 212, 214, 216,218, 220, 222, 234, 235, 237, 239, 243, 245, 247, 249, 251, 253, 255,265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291,293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319,321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347,349, 353, 355, 357, 359, 361, 363, 365, 367, 377, 379, 381, 383, 385,387, 389, 391, 393, 395, 397, 399, 406-419, 434-442, 448, 449, 462, 463or 464.

The present invention also provides an antibody or antigen-bindingfragment thereof that specifically binds LAG3 (e.g., human orcynomolgous monkey) comprising (1) a light chain variable domaincomprising CDR-L1 that comprises the amino acid sequence:KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprises the amino acidsequence: GASNLES (SEQ ID NO: 39); and CDR-L3 that comprises the aminoacid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variabledomain comprising: CDR-H1 that comprises the amino acid sequence: DYNVD(SEQ ID NO: 33); CDR-H2 that comprises the amino acid sequence:DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and CDR-H3 that comprises the aminoacid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (2) a light chain variabledomain comprising CDR-L1 that comprises the amino acid sequence:KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprises the amino acidsequence: GASNLES (SEQ ID NO: 39); and CDR-L3 that comprises the aminoacid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variabledomain comprising CDR-H1 that comprises the amino acid sequence: DYNVD(SEQ ID NO: 33); CDR-H2 that comprises the amino acid sequence:DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and CDR-H3 that comprises the aminoacid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (3) a light chain variabledomain comprising: CDR-L1 that comprises the amino acid sequence:KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprises the amino acidsequence: GASNLES (SEQ ID NO: 39); and CDR-L3 that comprises the aminoacid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variabledomain comprising CDR-H1 that comprises the amino acid sequence: DYNVD(SEQ ID NO: 33); CDR-H2 that comprises the amino acid sequence:DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3 that comprises the aminoacid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (4) a light chain variabledomain comprising CDR-L1 that comprises the amino acid sequence:KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprises the amino acidsequence: GASNLES (SEQ ID NO: 39); and CDR-L3 that comprises the aminoacid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variabledomain comprising: CDR-H1 that comprises the amino acid sequence: DYNVD(SEQ ID NO: 33); CDR-H2 that comprises the amino acid sequence:DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and CDR-H3 that comprises the aminoacid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (5) a light chain variabledomain comprising CDR-L1 that comprises the amino acid sequence:KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprises the amino acidsequence: GASNLES (SEQ ID NO: 39); and CDR-L3 that comprises the aminoacid sequence: QQSTEDPRT (SEQ ID NO: 40); and/or a heavy chain variabledomain comprising: CDR-H1 that comprises the amino acid sequence: DYNVD(SEQ ID NO: 33); CDR-H2 that comprises the amino acid sequence:DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3 that comprises the aminoacid sequence: NYRWFGAMDH (SEQ ID NO: 35).

In an embodiment of the invention, the antibody or antigen-bindingfragment thereof that specifically binds LAG3 (e.g., human orcynomolgous monkey LAG3) comprises the CDRs of various light and/orheavy chain variable regions and having at least 90% overall amino acidsequence identity to the variable region, i.e., variability in the chainoccurs outside the CDRs, e.g., (1) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:106 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 106or amino acids 1-119 thereof; (2) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:108 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 108or amino acids 1-119 thereof; (3) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:110 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 110or amino acids 1-119 thereof; (4) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:112 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 112or amino acids 1-119 thereof; (5) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:114 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 114or amino acids 1-119 thereof; (6) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:116 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 116or amino acids 1-119 thereof; (7) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:118 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 118or amino acids 1-119 thereof; (8) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:120 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 120or amino acids 1-119 thereof; and/or (9) a light chain immunoglobulincomprising CDR-L1, CDR-L2 and CDR-L3 of the immunoglobulin comprisingthe amino acid sequence of SEQ ID NO: 126 or amino acids 21-131 thereof,and having at least 90% amino acid sequence identity to the amino acidsequence set forth in SEQ ID NO: 126 or amino acids 21-131 thereof,and/or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3of the immunoglobulin comprising the amino acid sequence of SEQ ID NO:122 or amino acids 1-119 thereof, and having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 122or amino acids 1-119 thereof. In an embodiment of the invention, theantibody or antigen-binding fragment comprises the various mature lightand/or mature heavy chain immunoglobulin variable regions and having atleast 90% overall amino acid sequence identity to the unprocessedimmunoglobulin variable region (including the signal sequence), i.e.,variability occurs outside the mature immunoglobulin chain sequences.

The present invention provides an antibody or antigen-binding fragmentthereof that specifically binds LAG3 (e.g., human or cynomolgous monkeyLAG3) that comprises a mature or unprocessed V_(L) domain or light chainimmunoglobulin of SEQ ID NO: 7, 17, 27, 37, 57, 59, 61, 63, 65, 101,126, 130, 132, 136, 138, 208, 210, 224, 226, 228, 230, 232, 241, 257,259, 261, 263, 351, 369, 371, 373, 375, 401, 403, 405, 426, 427, 451-453or 459-461; and/or a mature or unprocessed V_(H) domain or heavy chainimmunoglobulin of SEQ ID NO: 2, 12, 22, 32, 45, 47, 49, 51, 53, 55, 67,69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 103,106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 128, 134, 140, 142,144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170,172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198,200, 202, 204, 206, 212, 214, 216, 218, 220, 222, 234, 235, 237, 239,243, 245, 247, 249, 251, 253, 255, 265, 267, 269, 271, 273, 275, 277,279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333,335, 337, 339, 341, 343, 345, 347, 349, 353, 355, 357, 359, 361, 363,365, 367, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399,406-419, 434-442, 448, 449, 462, 463 or 464. In an embodiment of theinvention, the antibody or antigen-binding fragment thereof comprises:(1) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40);and/or a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or(2) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40);and/or a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or(3) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40);and/or a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or(4) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40);and/or a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or(5) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40);and/or a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35). Inan embodiment of the invention, the antibody or fragment is glycosylatedwith engineered yeast N-linked glycans or CHO N-linked glycans.Optionally, any of the anti-LAG3 antibodies or antigen-binding fragmentsthereof of the invention are characterized by one or more of thefollowing properties: Inhibits LAG3 binding to MHC class II molecules;Competes with MHC class II molecules for LAG3 binding; Binds theextraloop of LAG3; Binds LAG3 with a K_(D) of about 10⁻⁹M to about2×10⁻¹²M; Binds to native LAG3 on the surface of activated CD4+ and/orCD8+ T-cells; Binds to human and/or cynomolgous monkey LAG3; InhibitsLAG3 homodimerization; Stimulates antigen-specific T-cell production ofIL-2; labels tonsil tissue; does not label brain, heart, kidney, liver,lung, pancreas, and/or pituitary tissue; binds to human LAG3 bycontacting residues QEGAPAQL (amino acids 35-42 of SEQ ID NO: 443) andRPARRADAGEYRAAVH (amino acids 137-152 of SEQ ID NO: 443) and,optionally, residues DERGRQRGDFSLW (amino acids 123-135 of SEQ ID NO:443) of LAG3; or residues SPTIPLQDL (amino acids 45-53 of SEQ ID NO:443) and, optionally DERGRQRGDFSL (amino acids 123-134 of SEQ ID NO:443) of LAG3; or residues HPLAPGPHPAAPSSWGPRPRRYTVL (amino acids 78-102of SEQ ID NO: 443) of LAG3; and/or by protecting hydrogens on the amidebackbone of such residues from exchange with a deuterium. The presentinvention also provides any such antibody or fragment in apharmaceutically acceptable carrier or diluent. In an embodiment of theinvention, the anti-LAG3 antibody or fragment is immobilized to a solidsubstrate. In an embodiment of the invention the anti-LAG3 antibody orantigen-binding fragment thereof is Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 or Ab9 (as set forth herein).

The present invention also provides a complex comprising an anti-LAG3antibody or fragment discussed herein (e.g., 4A10, 19E8, 11C9 and/or22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) complexedwith LAG3 (e.g., human or cynomolgous monkey) or a fragment thereof orwith a secondary antibody (e.g., detectably labeled secondary antibody)that binds specifically to the anti-LAG3 antibody or fragment. In anembodiment of the invention, the antibody or fragment is in vitro (e.g.,is immobilized to a solid substrate) or is in the body of a subject. Inan embodiment of the invention, the LAG3 is in vitro (e.g., isimmobilized to a solid substrate) or is on the surface of a cell or isin the body of a subject.

The present invention further provides a composition comprising aplurality of anti-LAG3 antibodies or antigen-binding fragments of thepresent invention which are isolated, monoclonal antibodies orantigen-binding fragments thereof, e.g., which are humanized, e.g.,humanized antagonistic antibodies and antigen-binding fragments thereof.

The present invention also provides a composition comprising anti-LAG3antibodies or antigen-binding fragments thereof as discussed herein inassociation with a further therapeutic agent (e.g., a monoclonalantibody or antigen-binding fragment thereof or an organic smallmolecule) such as an inhibitor of an immunomodulatory receptor, ananti-emetic, an MTOR (mammalian target of rapamycin) inhibitor, acytotoxic agent, a platinum agent, an EGFR (epidermal growth factorreceptor) inhibitor, a VEGF (vascular epidermal growth factor)inhibitor, a microtubule stabilizer, a taxane, a CD20 inhibitor, a CD52inhibitor, a CD30 inhibitor, a RANK (Receptor activator of nuclearfactor kappa-B) inhibitor, a RANKL (Receptor activator of nuclear factorkappa-B ligand) inhibitor, an ERK inhibitor, a MAP Kinase inhibitor, anAKT inhibitor, a MEK inhibitor, a PI3K inhibitor, a HER1 inhibitor, aHER2 inhibitor, a HER3 inhibitor, a HER4 inhibitor, a Bcl2 inhibitor, aCD22 inhibitor, a CD79b inhibitor, an ErbB2 inhibitor, and/or a farnesylprotein transferase inhibitor. In an embodiment of the invention, thefurther therapeutic agent is an anti-PD1 antibody or an antigen-bindingfragment thereof, pembrolizumab, nivolumab, CT-011, anti-CTLA4 antibodyor an antigen-binding fragment thereof, anti-TIM3 antibody or anantigen-binding fragment thereof, anti-CS1 antibody or anantigen-binding fragment thereof, elotuzumab, anti-KIR2DL1/2/3 antibodyor an antigen-binding fragment thereof, lirilumab, anti-CD137 antibodyor an antigen-binding fragment thereof, urelumab, anti-GITR antibody oran antigen-binding fragment thereof, TRX518, anti-PD-L1 antibody or anantigen-binding fragment thereof, BMS-936559, MSB0010718C, MPDL3280A,anti-PD-L2 antibody or an antigen-binding fragment thereof, anti-ILT1antibody or an antigen-binding fragment thereof, anti-ILT2 antibody oran antigen-binding fragment thereof, anti-ILT3 antibody or anantigen-binding fragment thereof, anti-ILT4 antibody or anantigen-binding fragment thereof, anti-ILT5 antibody or anantigen-binding fragment thereof, anti-ILT6 antibody or anantigen-binding fragment thereof, anti-ILT7 antibody or anantigen-binding fragment thereof, anti-ILT8 antibody or anantigen-binding fragment thereof, anti-CD40 antibody or anantigen-binding fragment thereof, anti-OX40 antibody or anantigen-binding fragment thereof, anti-CD137 antibody or anantigen-binding fragment thereof, anti-KIR2DL1 antibody or anantigen-binding fragment thereof, anti-KIR2DL2/3 antibody or anantigen-binding fragment thereof, anti-KIR2DL4 antibody or anantigen-binding fragment thereof, anti-KIR2DL5A antibody or anantigen-binding fragment thereof, anti-KIR2DL5B antibody or anantigen-binding fragment thereof, anti-KIR3DL1 antibody or anantigen-binding fragment thereof, anti-KIR3DL2 antibody or anantigen-binding fragment thereof, anti-KIR3DL3 antibody or anantigen-binding fragment thereof, anti-NKG2A antibody or anantigen-binding fragment thereof, anti-NKG2C antibody or anantigen-binding fragment thereof, and/or an anti-NKG2E antibody or anantigen-binding fragment thereof, or any small organic moleculeinhibitor of such targets; IL-10, anti-IL10, anti-TSLP and/or PEGylatedIL-10. In an embodiment of the invention, the further therapeutic agentis 13-cis-retinoic acid,3-[5-(methylsulfonylpiperadinemethyl)-indolyl]-quinolone,4-hydroxytamoxifen, 5-deooxyuridine, 5′-deoxy-5-fluorouridine,5-fluorouracil, 6-mecaptopurine, 7-hydroxystaurosporine, A-443654,abirateroneacetate, abraxane, ABT-578, acolbifene, ADS-100380,aflibercept, ALT-110, altretamine, amifostine, aminoglutethimide,amrubicin, amsacrine, anagrelide, anastrozole, angiostatin, AP-23573,ARQ-197, arzoxifene, AS-252424, AS-605240, asparaginase, AT-9263,ATI3387, atrasentan, axitinib, AZD1152, Bacillus Calmette-Guerin (BCG)vaccine, batabulin, BC-210, BGJ398, besodutox, bevacizumab,bicalutamide, Bio111, BIO140, BKM120, bleomycin, BMS-214662, BMS-247550,BMS-275291, BMS-310705, bortezimib, buserelin, busulfan, calcitriol,camptothecin, canertinib, capecitabine, carboplatin, carmustine, CC8490,cediranib, CG-1521, CG-781, chlamydocin, chlorambucil, chlorotoxin,cilengitide, cimitidine, cisplatin, cladribine, clodronate, cobimetnib,COL-3, CP-724714, cyclophosphamide, cyproterone, cyproteroneacetate,cytarabine, cytosinearabinoside, dabrafenib, dacarbazine, dacinostat,dactinomycin, dalotuzumab, danusertib, dasatanib, daunorubicin,decatanib, deguelin, denileukin, deoxycoformycin, depsipeptide,diarylpropionitrile, diethylstilbestrol, diftitox, DNE03, docetaxel,dovitinib, doxorubicin, droloxifene, edotecarin, yttrium-90labeled-edotreotide, edotreotide, EKB-569, EMD121974, encorafenib,endostatin, enzalutamide, enzastaurin, epirubicin, epithilone B,ERA-923, erbitux, erlotinib, estradiol, estramustine, etoposide,everolimus, exemestane, ficlatuzumab, finasteride, flavopiridol,floxuridine, fludarabine, fludrocortisone, fluoxymesterone, flutamide,FOLFOX regimen, fulvestrant, galeterone, ganetespib, gefitinib,gemcitabine, gimatecan, goserelin, goserelin acetate, gossypol,GSK461364, GSK690693, HMR-3339, hydroxyprogesteronecaproate,hydroxyurea, IC87114, idarubicin, idoxyfene, ifosfamide, IM862,imatinib, IMC-1C11, INCB24360, INC280, INO1001, interferon,interleukin-12, ipilimumab, irinotecan, JNJ-16241199, ketoconazole,KRX-0402, lapatinib, lasofoxifene, LEE011, letrozole, leucovorin,leuprolide, leuprolide acetate, levamisole, liposome entrappedpaclitaxel, lomustine, lonafarnib, lucanthone, LY292223, LY292696,LY293646, LY293684, LY294002, LY3009120, LY317615, marimastat,mechlorethamine, medroxyprogesteroneacetate, megestrolacetate, MEK162,melphalan, mercaptopurine, mesna, methotrexate, mithramycin, mitomycin,mitotane, mitoxantrone, tozasertib, MLN8054, natitoclax, neovastat,neratinib, neuradiab, nilotinib, nilutimide, nolatrexed, NVP-BEZ235,oblimersen, octreotide, ofatumumab, oregovomab, ornatuzumab, orteronel,oxaliplatin, paclitaxel, palbociclib, pamidronate, panitumumab,pazopanib, PD0325901, PD184352, PEG-interferon, pemetrexed, pentostatin,perifosine, phenylalaninemustard, PI-103, pictilisib, PIK-75,pipendoxifene, PKI-166, plicamycin, PLX8394, porfimer, prednisone,procarbazine, progestins, PX-866, R-763, raloxifene, raltitrexed,razoxin, ridaforolimus, rituximab, romidepsin, RTA744, rubitecan,scriptaid, Sdx102, seliciclib, selumetinib, semaxanib, SF1126,sirolimus, SN36093, sorafenib, spironolactone, squalamine, SR13668,streptozocin, SU6668, suberoylanalide hydroxamic acid, sunitinib,synthetic estrogen, talampanel, talimogene laherparepvec, tamoxifen,temozolomide, temsirolimus, teniposide, tesmilifene, testosterone,tetrandrine, TGX-221, thalidomide, 6-thioguanine, thiotepa, ticilimumab,tipifarnib, tivozanib, TKI-258, TLK286, topotecan, toremifene citrate,trabectedin, trametinib, trastuzumab, tretinoin, trichostatin A,triciribinephosphate monohydrate, triptorelin pamoate, TSE-424, uracilmustard, valproic acid, valrubicin, vandetanib, vatalanib, VEGF trap,vemurafenib, vinblastine, vincristine, vindesine, vinorelbine, vitaxin,vitespan, vorinostat, VX-745, wortmannin, Xr311, zanolimumab, ZK186619,ZK-304709, ZM336372, ZSTK474, casopitant, netupitant, palonosetron,aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam,alprazolam, haloperidol, droperidol, dronabinol, dexamethasone,methylprednisolone, prochlorperazine, granisetron, ondansetron,dolasetron, tropisetron, GCSF, PEG-GCSF, erythropoietin, epoetin alfaand darbepoetin alfa. In an embodiment of the invention, the anti-LAG3antibody or antigen-binding fragment thereof is in association withpembrolizumab. In an embodiment of the invention, the anti-LAG3 antibodyor antigen-binding fragment thereof is in association with novolumab. Inan embodiment of the invention, the anti-LAG3 antibody orantigen-binding fragment thereof is in association with CT-011. In anembodiment of the invention, the anti-LAG3 antibody or antigen-bindingfragment thereof is in association with BMS-936559.

The present invention also provides a polypeptide comprising an aminoacid sequence selected from the group consisting of 2, 3, 4, 5, 7, 8, 9,10, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 27, 28, 29, 30, 32,33, 34, 35, 37, 38, 39, 40, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65,67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101,103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130,132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158,160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186,188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214,216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 237, 239, 241,243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269,271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297,299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325,327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353,355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381,383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 406, 407,408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 426, 427,434, 435, 436, 437, 438, 439, 440, 441, 442, 446, 448, 449, 451, 452,453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463 and 464 forexample, SEQ ID NOs: 2, 7, 12, 17, 22, 27, 32 and 37; or a maturefragment thereof. The present invention also provides a polynucleotideencoding any of such polypeptides, e.g., comprising a nucleotidesequence selected from the group consisting of SEQ ID NOs: 1, 6, 11, 16,21, 26, 31, 36, 46, 48, 50, 52, 54, 56, 56, 58, 60, 62, 64, 66, 68, 70,72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104,105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131,133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159,161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187,189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215,217, 219, 221, 223, 225, 227, 229, 231, 233, 236, 238, 240, 242, 244,246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272,274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300,302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328,330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356,358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384,386, 388, 390, 392, 394, 396, 398, 400, 402 and 404; or a maturefragment thereof. Also included in the present invention is a vector(e.g., a plasmid) comprising the polynucleotide. A host cell (e.g.,mammalian, bacterial, Chinese hamster ovary (CHO), lower eukaryotic,fungal, yeast, Pichia, Pichia pastoris) is also part of the presentinvention wherein the host cell comprises an antibody, fragment,polypeptide, polynucleotide and/or vector set forth herein.

The present invention also provides vaccines comprising an antibody orfragment set forth herein, an antigen (e.g., viral peptide antigen,virus-like particle, tumor peptide antigen) and a pharmaceuticallyacceptable carrier.

The prevent invention also provides a vessel (e.g., plastic or glassvial) or an injection device (e.g., a syringe such as a pre-filledsyringe or an autoinjector) comprising any antibody, fragment,polypeptide, polynucleotide, vector, composition or vaccine discussedherein.

The present invention also provides a method of treating a cancer (e.g.,osteosarcoma, rhabdomyosarcoma, neuroblastoma, kidney cancer, leukemia,renal transitional cell cancer, bladder cancer, Wilm's cancer, ovariancancer, pancreatic cancer, breast cancer, prostate cancer, bone cancer,lung cancer, gastric cancer, colorectal cancer, cervical cancer,synovial sarcoma, head and neck cancer, squamous cell carcinoma,multiple myeloma, renal cell cancer, retinoblastoma, hepatoblastoma,hepatocellular carcinoma, melanoma, rhabdoid tumor of the kidney,Ewing's sarcoma, chondrosarcoma, brain cancer, glioblastoma, meningioma,pituitary adenoma, vestibular schwannoma, a primitive neuroectodermaltumor, medulloblastoma, astrocytoma, anaplastic astrocytoma,oligodendroglioma, ependymoma, choroid plexus papilloma, polycythemiavera, thrombocythemia, idiopathic myelfibrosis, soft tissue sarcoma,thyroid cancer, endometrial cancer, carcinoid cancer or liver cancer,breast cancer or gastric cancer) in a subject, comprising administeringto the subject a effective amount of an anti-LAG3 antibody orantigen-binding fragment or vaccine discussed herein optionally, inassociation with a further therapeutic agent or a therapeutic procedure(e.g., surgical tumorectomy or anti-cancer radiation therapy).

The present invention also provides a method of administering ananti-LAG3 antibody, fragment, composition, polypeptide, vaccine orpolynucleotide discussed herein, or a pharmaceutical compositionthereof, to a subject comprising injecting the antibody, fragment,polypeptide, vaccine or polynucleotide into the body of the subjectusing an injection device; and, optionally, also administering a furthertherapeutic agent to the subject.

The present invention also provides a method of producing an anti-LAG3antibody or antigen-binding fragment thereof or polypeptide discussedherein comprising: a. culturing a host cell comprising a polynucleotideencoding the polypeptide or an immunoglobulin chain of the antibody orfragment in culture medium under conditions favorable to expression ofthe polynucleotide; and b. optionally, recovering the antibody, fragmentor polypeptide from the host cell and/or culture medium. In anembodiment of the invention, the method comprises the step ofintroducing the polynucleotide into the host cell, e.g., bytransformation or transfection.

The present invention also provides a method for detecting the presenceof a LAG3 peptide or a fragment thereof in a sample comprisingcontacting the sample with an anti-LAG3 antibody or fragment discussedherein and detecting the presence of a complex between the antibody orfragment and the peptide; wherein detection of the complex indicates thepresence of the LAG3 peptide.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Binding of anti-human LAG3 clones to human and cynomolgus monkeyLAG3 expressing CHO-K1 cells.

FIG. 2. Binding of LAG3 clones to cynomolgous monkey and human primaryT-cells.

FIG. 3. Three-dimensional structure of human LAG3. The location of theantibody 22D2, 11C9 and 4A10 epitopes are indicated.

FIG. 4 (a-c 1). Heat map indicating regions in human LAG3 which arestrongly or weakly protected from deuteration by antibody binding. (a)and (a-1) human LAG3/22D2 Difference heatmap; (b) and (b-1) humanLAG3/11C9 Difference heatmap; (c) and (c-1) human LAG3/4A10 Differenceheatmap.

FIG. 5(a-b). Combined heatmaps indicating (a) and (a-1) regions on humanLAG3 which are protected from deuteration by 22D2, 11C9 and 4A10 bindingand (b) the location in human LAG3 mediating MHC2 binding.

FIG. 6. Predominant N-linked glycans for monoclonal antibodies producedin Chinese hamster ovary cells (CHO N-linked glycans) and in engineeredyeast cells (engineered yeast N-linked glycans): squares:N-acetylglucosamine (GlcNac); circles: mannose (Man); diamonds:galactose (Gal); triangles: fucose (Fuc).

FIG. 7. Effect of anti-human LAG-3 antibody treatment+/−anti-PD-1treatment on IL-2 production in SEB stimulated human PBMCs. PBMC wereactivated by SEB for 3 days and IL-2 concentration in culturesupernatants was determined by MSD. Donor 090: SEB 60 ng/mL, Donor 089:SEB 30 ng/mL. Anti-PD1 was used at 10 μg/mL.

FIG. 8. Effect of hu22D2 treatment+/−anti-PD-1 treatment on IFN-γ andTNFα production in MLR stimulated human PBMCs. PBMC were activated withallogeneic monocyte-derived DC for 7 days and IFN-γ and TNF-αconcentration in culture supernatants was determined by MSD. Anti-PD-1was used at 3 μg/mL. Isotype was used at 200 nM.

FIG. 9. Antigen-capture assay analysis of concentration of unbound andpartially bound anti-LAG3 antibody Ab6 in cynomolgous monkey subjectsover time at various doses (0.03 mg/kg; 0.3 mg/kg; 1 mg/kg; 10 mg/kg; 30mg/kg).

FIG. 10. Universal assay analysis of concentration of total anti-LAG3antibody Ab6 in cynomolgous monkey subjects over time at various doses(0.03 mg/kg; 0.3 mg/kg; 1 mg/kg; 10 mg/kg; 30 mg/kg).

FIG. 11. The dose-normalized concentration data of Ab6 over time incynomolgous monkey subjects at various doses (0.03 mg/kg; 0.3 mg/kg; 1mg/kg; 10 mg/kg; 30 mg/kg).

FIG. 12. Target related clearance (Vmax, Km) evaluation of various doesof anti-LAG3 antibody Ab6 at various doses.

DETAILED DESCRIPTION

The present invention provides antibodies and antigen-binding fragmentsthereof that have exceptionally high affinity for human LAG3 andcynomolgous monkey LAG3, e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 orAb9 as well as uses thereof and methods of making the same as isdiscussed herein. For example, affinity (K_(D)) for human LAG3 by KinExAassay was measured to be as high as 2 pM and affinity for cynomolgousLAG3 is in the low double digit pM range. A particularly low isoelectricpoint (e.g., about 6.3) makes some of these antibodies unique. Moreover,though some antibodies bind LAG3 primarily outside the extraloop region,they exhibit the ability to block LAG3/MCH class II binding.Furthermore, the anti-LAG3 antibodies of the present invention have ahigh degree of specificity for binding to LAG3 over other relatedproteins. Such antibodies and fragments set forth herein may be useful,for example, for treatment of various cancers and infectious diseases.

Abbreviations

Throughout the detailed description and examples of the invention thefollowing abbreviations will be used:

-   ADCC Antibody-dependent cellular cytotoxicity-   CDC Complement-dependent cytotoxicity-   CDR Complementarity determining region in the immunoglobulin    variable regions, defined using the Kabat numbering system-   CHO Chinese hamster ovary-   EC50 concentration resulting in 50% efficacy or binding-   ELISA Enzyme-linked immunosorbant assay-   FR Antibody framework region: the immunoglobulin variable regions    excluding the CDR regions.-   HRP Horseradish peroxidase-   IC50 concentration resulting in 50% inhibition-   IgG Immunoglobulin G-   Kabat An immunoglobulin alignment and numbering system pioneered by    Elvin A. Kabat ((1991) Sequences of Proteins of Immunological    Interest, 5th Ed. Public Health Service, National Institutes of    Health, Bethesda, Md.)-   mAb or Mab or MAb Monoclonal antibody-   PCR Polymerase chain reaction-   V region The segment of IgG chains which is variable in sequence    between different antibodies. It extends to Kabat residue 109 in the    light chain and 113 in the heavy chain.-   VH Immunoglobulin heavy chain variable region-   VK Immunoglobulin kappa light chain variable region

Definitions

So that the invention may be more readily understood, certain technicaland scientific terms are specifically defined below. Unless specificallydefined elsewhere in this document, all other technical and scientificterms used herein have the meaning commonly understood by one ofordinary skill in the art to which this invention belongs.

As used herein, including the appended claims, the singular forms ofwords such as “a,” “an,” and “the,” include their corresponding pluralreferences unless the context clearly dictates otherwise.

LAG3

The term “LAG3”, with respect to the polypeptide to which antibodies andantigen-binding fragments of the present invention bind, refers to humanand cynomolgous monkey, e.g., Macaca fascicularis or Macaca mulatta LAG3as well as fragments thereof such as the mature fragment thereof lackingthe signal peptide.

In an embodiment of the invention, the amino acid sequence of human LAG3(Lymphocyte Activation Gene-3) comprises the amino acid sequence:

(SEQ ID NO: 443) MWEAQFLGLL FLQPLWVAPV KPLQPGAEVPVVWAQEGAPA QLPCSPTIPL QDLSLLRRAG VTWQHQPDSG PPAAAPGHPL APGPHPAAPSSWGPRPRRYT VLSVGPGGLR SGRLPLQPRV QLDERGRQRG DFSLWLRPAR RADAGEYRAAVHLRDRALSC RLRLRLGQAS MTASPPGSLR ASDWVILNCS FSRPDRPASV HWFRNRGQGRVPVRESPHHH LAESFLFLPQ VSPMDSGPWG CILTYRDGFN VSIMYNLTVL GLEPPTPLTVYAGAGSRVGL PCRLPAGVGT RSFLTAKWTP PGGGPDLLVT GDNGDFTLRL EDVSQAQAGTYTCHIHLQEQ QLNATVTLAI ITVTPKSFGS PGSLGKLLCE VTPVSGQERF VWSSLDTPSQRSFSGPWLEA QEAQLLSQPW QCQLYQGERL LGAAVYFTEL SSPGAQRSGR APGALPAGHLLLFLILGVLS LLLLVTGAFG FHLWRRQWRP RRFSALEQGI HPPQAQSKIE ELEQEPEPEPEPEPEPEPEP EPEQL;see also Uniprot accession no. P18627.

In an embodiment of the invention, the amino acid sequence of mouse LAG3comprises the amino acid sequence:

(SEQ ID NO: 444) MREDLLLGFL LLGLLWEAPV VSSGPGKELPVVWAQEGAPV HLPCSLKSPN LDPNFLRRGG VIWQHQPDSG QPTPIPALDL HQGMPSPRQPAPGRYTVLSV APGGLRSGRQ PLHPHVQLEE RGLQRGDFSL WLRPALRTDA GEYHATVRLPNRALSCSLRL RVGQASMIAS PSGVLKLSDW VLLNCSFSRP DRPVSVHWFQ GQNRVPVYNSPRHFLAETFL LLPQVSPLDS GTWGCVLTYR DGFNVSITYN LKVLGLEPVA PLTVYAAEGSRVELPCHLPP GVGTPSLLIA KWTPPGGGPE LPVAGKSGNF TLHLEAVGLA QAGTYTCSIHLQGQQLNATV TLAVITVTPK SFGLPGSRGK LLCEVTPASG KERFVWRPLN NLSRSCPGPVLEIQEARLLA ERWQCQLYEG QRLLGATVYA AESSSGAHSA RRISGDLKGG HLVLVLILGALSLFLLVAGA FGFHWWRKQL LLRRFSALEH GIQPFPAQRK IEELERELET EMGQEPEPEPEPQLEPEPRQ L;See also Uniprot accession no. Q61790

In an embodiment of the invention, the amino acid sequence ofcynomolgous monkey LAG3 comprises the amino acid sequence:

(SEQ ID NO: 445) MWEAQFLGLL FLQPLWVAPV KPPQPGAEISVVWAQEGAPA QLPCSPTIPL QDLSLLRRAG VTWQHQPDSG PPAXAPGHPP VPGHRPAAPYSWGPRPRRYT VLSVGPGGLR SGRLPLQPRV QLDERGRQRG DFSLWLRPAR RADAGEYRATVHLRDRALSC RLRLRVGQAS MTASPPGSLR TSDWVILNCS FSRPDRPASV HWFRSRGQGRVPVQGSPHHH LAESFLFLPH VGPMDSGLWG CILTYRDGFN VSIMYNLTVL GLEPATPLTVYAGAGSRVEL PCRLPPAVGT QSFLTAKWAP PGGGPDLLVA GDNGDFTLRL EDVSQAQAGTYICHIRLQGQ QLNATVTLAI ITVTPKSFGS PGSLGKLLCE VTPASGQEHF VWSPLNTPSQRSFSGPWLEA QEAQLLSQPW QCQLHQGERL LGAAVYFTEL SSPGAQRSGR APGALRAGHLPLFLILGVLF LLLLVTGAFG FHLWRRQWRP RRFSALEQGI HPPQAQSKIE ELEQEPELEPEPELERELGP EPEPGPEPEP EQL;See also NCBI reference number XP_005570011.1

The mature sequence of human, mouse and cynomolgous monkey LAG3, i.e.the sequence after removal of the signal peptide, comprises amino acids1-28 of SEQ ID NO: 443, 444 or 445.

LAG3 sequences may differ, for example, by having, for example,conserved mutations or mutations in non-conserved regions, e.g., whereinthe LAG3 has substantially the same biological function as the LAG3 ofSEQ ID NOs: 443 or 445. For example, biological functions of LAG3 are tobind to major histocompatibility complex (MHC) class II molecules and toform homodimers.

A particular LAG3 sequence will generally be at least 90% identical inamino acid sequence to LAG3 of SEQ ID NOs: 443 or 445 or other isoforms.In certain cases, a LAG3 may be at least 95%, or even at least 96%, 97%,98% or 99% identical to a LAG3 of SEQ ID NOs: 443 or 445, or otherisoforms or variants. In certain embodiments, a LAG3 sequence willdisplay no more than 10 amino acid differences from the LAG3 of any ofSEQ ID NOs: 443 or 445, or other isoforms or variants. In certainembodiments, the LAG3 may display no more than 5, or even no more than4, 3, 2, or 1 amino acid difference from a LAG3 of SEQ ID NOs: 443 or445, or other isoforms or variants. Percent identity can be determinedas described herein.

Anti-LAG3 Antibodies and Antigen-Binding Fragments Thereof

The present invention provides antibodies or antigen-binding fragmentsthereof that specifically bind LAG3 (e.g., human and/or cynomolgousmonkey, e.g., Macaca fascicularis or Macaca mulatta LAG3), for example,4A10, 19E8, 11C9, 22D2, e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 orAb9, and uses of such antibodies or fragments. In an embodiment of theinvention, the antibody or fragment is an antibody.

As used herein, an anti-LAG3 antibody or antigen-binding fragmentthereof refers to an antibody or antigen-binding fragment thereof thatspecifically binds to human or cynomolgous monkey LAG3. An antibodybinds specifically to a polypeptide comprising a given sequence (in thiscase an epitope of human or cynomolgous monkey LAG3) if it binds topolypeptides comprising the LAG3 sequence with a K_(D) of about 1 nM ora higher affinity (e.g., 1 nM-2 pM, 1 nM, 100 pM, 10 pM or 2 pM), butdoes not bind to proteins lacking the sequence. For example, an antibodyor antigen-binding fragment that specifically binds to a polypeptidecomprising human or cynomolgous monkey LAG3 may bind to a FLAG®-taggedform of human or cynomolgous monkey LAG3 but will not bind to otherFLAG®-tagged proteins that lack LAG3 epitopes.

The present invention includes anti-LAG3 antibodies and methods of usethereof. As used herein, the term “antibody” refers to any form ofantibody that exhibits the desired biological activity. Thus, it is usedin the broadest sense and specifically covers, but is not limited to,monoclonal antibodies (including full length monoclonal antibodies),polyclonal antibodies, multispecific antibodies (e.g., bispecificantibodies), humanized antibodies, fully human antibodies, chimericantibodies and camelized single domain antibodies.

The present invention includes parental anti-LAG3 antibodies andantigen-binding fragments thereof and methods of use thereof “Parentalantibodies and antigen-binding fragments thereof” are antibodies andfragments which may be modified for an intended use, such ashumanization of an antibody for use as a human therapeutic antibody orfragment.

The present invention includes anti-LAG3 antigen-binding fragments andmethods of use thereof. As used herein, unless otherwise indicated,“antibody fragment” or “antigen-binding fragment” refers toantigen-binding fragments of antibodies, i.e. antibody fragments thatretain the ability to bind specifically to the antigen bound by thefull-length antibody, e.g. fragments that retain one or more CDRregions. Examples of antigen-binding fragments include, but are notlimited to, Fab, Fab′, F(ab′)₂, and Fv fragments; diabodies;single-chain antibody molecules, e.g., sc-Fv; nanobodies andmultispecific antibodies formed from antibody fragments.

The present invention includes anti-LAG3 Fab fragments and methods ofuse thereof. A “Fab fragment” is comprised of one light chain and theC_(H)1 and variable regions of one heavy chain. The heavy chain of a Fabmolecule cannot form a disulfide bond with another heavy chain molecule.An “Fab fragment” can be the product of papain cleavage of an antibody.

The present invention includes anti-LAG3 antibodies and antigen-bindingfragments thereof which comprise an Fc region and methods of usethereof. An “Fc” region contains two heavy chain fragments comprisingthe C_(H)1 and C_(H)2 domains of an antibody. The two heavy chainfragments are held together by two or more disulfide bonds and byhydrophobic interactions of the C_(H)3 domains.

The present invention includes anti-LAG3 Fab′ fragments and methods ofuse thereof. A “Fab′ fragment” contains one light chain and a portion orfragment of one heavy chain that contains the V_(H) domain and theC_(H)1 domain and also the region between the C_(H)1 and C_(H)2 domains,such that an interchain disulfide bond can be formed between the twoheavy chains of two Fab′ fragments to form a F(ab′)₂ molecule.

The present invention includes anti-LAG3 F(ab′)₂ fragments and methodsof use thereof. A “F(ab′)₂ fragment” contains two light chains and twoheavy chains containing a portion of the constant region between theC_(H1) and C_(H2) domains, such that an interchain disulfide bond isformed between the two heavy chains. A F(ab′)₂ fragment thus is composedof two Fab′ fragments that are held together by a disulfide bond betweenthe two heavy chains. An “F(ab′)₂ fragment” can be the product of pepsincleavage of an antibody.

The present invention includes anti-LAG3 Fv fragments and methods of usethereof. The “Fv region” comprises the variable regions from both theheavy and light chains, but lacks the constant regions.

The present invention includes anti-LAG3 scFv fragments and methods ofuse thereof. The term “single-chain Fv” or “scFv” antibody refers toantibody fragments comprising the V_(H) and V_(L) domains of anantibody, wherein these domains are present in a single polypeptidechain. Generally, the Fv polypeptide further comprises a polypeptidelinker between the V_(H) and V_(L) domains which enables the scFv toform the desired structure for antigen-binding. For a review of scFv,see Pluckthun (1994) THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol.113, Rosenburg and Moore eds. Springer-Verlag, N.Y., pp. 269-315. Seealso, International Patent Application Publication No. WO88/01649 andU.S. Pat. Nos. 4,946,778 and 5,260,203.

The present invention includes anti-LAG3 domain antibodies and methodsof use thereof. A “domain antibody” is an immunologically functionalimmunoglobulin fragment containing only the variable region of a heavychain or the variable region of a light chain. In some instances, two ormore V_(H) regions are covalently joined with a peptide linker to createa bivalent domain antibody. The two V_(H) regions of a bivalent domainantibody may target the same or different antigens.

The present invention includes anti-LAG3 bivalent antibodies and methodsof use thereof. A “bivalent antibody” comprises two antigen-bindingsites. In some instances, the two binding sites have the same antigenspecificities. However, bivalent antibodies may be bispecific (seebelow).

The present invention includes anti-LAG3 camelized single domainantibodies and methods of use thereof. In certain embodiments,antibodies herein also include camelized single domain antibodies. See,e.g., Muyldermans et al. (2001) Trends Biochem. Sci. 26:230; Reichmannet al. (1999) J. Immunol. Methods 231:25; WO 94/04678; WO 94/25591; U.S.Pat. No. 6,005,079). In one embodiment, the present invention providessingle domain antibodies comprising two V_(H) domains with modificationssuch that single domain antibodies are formed.

The present invention includes anti-LAG3 diabodies and methods of usethereof. As used herein, the term “diabodies” refers to small antibodyfragments with two antigen-binding sites, which fragments comprise aheavy chain variable domain (V_(H)) connected to a light chain variabledomain (V_(L)) in the same polypeptide chain (V_(H)-V_(L) orV_(L)-V_(H)). By using a linker that is too short to allow pairingbetween the two domains on the same chain, the domains are forced topair with the complementary domains of another chain and create twoantigen-binding sites. Diabodies are described more fully in, e.g., EP404,097; WO 93/11161; and Holliger et al. (1993) Proc. Natl. Acad. Sci.USA 90: 6444-6448. For a review of engineered antibody variantsgenerally see Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

Typically, an antibody or antigen-binding fragment of the inventionwhich is modified in some way retains at least 10% of its LAG3 (e.g.,human and/or cynomolgous monkey, e.g., Macaca fascicularis or Macacamulatta LAG3) binding activity (when compared to the parental antibody)when that activity is expressed on a molar basis. Preferably, anantibody or antigen-binding fragment of the invention retains at least20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the LAG3 (e.g., humanand/or cynomolgous monkey, e.g., Macaca fascicularis or Macaca mulattaLAG3) binding affinity as the parental antibody. It is also intendedthat an antibody or antigen-binding fragment of the invention caninclude conservative or non-conservative amino acid substitutions(referred to as “conservative variants” or “function conserved variants”of the antibody) that do not substantially alter its biologic activity.

The present invention includes isolated anti-LAG3 antibodies andantigen-binding fragments thereof (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) and methods ofuse thereof as well as isolated polypeptide immunoglobulin chainsthereof and isolated polynucleotides encoding such polypeptides andisolated vectors including such polynucleotides. “Isolated” antibodiesor antigen-binding fragments thereof, polypepotides, polynucleotides andvectors, are at least partially free of other biological molecules fromthe cells or cell culture from which they are produced. Such biologicalmolecules include nucleic acids, proteins, lipids, carbohydrates, orother material such as cellular debris and growth medium. An isolatedantibody or antigen-binding fragment may further be at least partiallyfree of expression system components such as biological molecules from ahost cell or of the growth medium thereof. Generally, the term“isolated” is not intended to refer to a complete absence of suchbiological molecules or to an absence of water, buffers, or salts or tocomponents of a pharmaceutical formulation that includes the antibodiesor fragments.

The present invention includes monoclonal anti-LAG3 antibodies andantigen-binding fragments thereof as well as monoclonal compositionscomprising a plurality of isolated monoclonal antibodies. The term“monoclonal antibody”, as used herein, refers to a population ofsubstantially homogeneous antibodies, i.e., the antibody moleculescomprising the population are identical in amino acid sequence exceptfor possible naturally occurring mutations that may be present in minoramounts. A “plurality” of such monoclonal antibodies and fragments in acomposition refers to a concentration of identical (i.e., as discussedabove, in amino acid sequence except for possible naturally occurringmutations that may be present in minor amounts) antibodies and fragmentswhich is above that which would normally occur in nature, e.g., in theblood of a host organism such as a mouse or a human. In contrast,conventional (polyclonal) antibody preparations typically include amultitude of different antibodies having different amino acid sequencesin their variable domains, particularly their CDRs, that are oftenspecific for different epitopes. The modifier “monoclonal” indicates thecharacter of the antibody as being obtained from a substantiallyhomogeneous population of antibodies, and is not to be construed asrequiring production of the antibody by any particular method. Forexample, the monoclonal antibodies to be used in accordance with thepresent invention may be made by the hybridoma method first described byKohler et al. (1975) Nature 256: 495, or may be made by recombinant DNAmethods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonalantibodies” may also be isolated from phage antibody libraries using thetechniques described in Clackson et al. (1991) Nature 352: 624-628 andMarks et al. (1991) J. Mol. Biol. 222: 581-597, for example. See alsoPresta (2005) J. Allergy Clin. Immunol. 116:731.

The present invention includes anti-LAG3 chimeric antibodies (e.g.,human constant domain/mouse variable domain) and methods of use thereof.As used herein, a “chimeric antibody” is an antibody having the variabledomain from a first antibody and the constant domain from a secondantibody, where the first and second antibodies are from differentspecies. (U.S. Pat. No. 4,816,567; and Morrison et al., (1984) Proc.Natl. Acad. Sci. USA 81: 6851-6855). Typically, the variable domains areobtained from an antibody from an experimental animal (the “parentalantibody”), such as a rodent, and the constant domain sequences areobtained from human antibodies, so that the resulting chimeric antibodywill be less likely to elicit an adverse immune response in a humansubject than the parental (e.g., mouse) antibody.

The present invention includes anti-LAG3 humanized antibodies andantigen-binding fragments thereof (e.g., mouse antibodies that have beenhumanized) and methods of use thereof. As used herein, the term“humanized antibody” refers to forms of antibodies that containsequences from both human and non-human (e.g., mouse or rat) antibodies.In general, the humanized antibody will comprise substantially all of atleast one, and typically two, variable domains, in which all orsubstantially all of the hypervariable loops correspond to those of anon-human immunoglobulin, and all or substantially all of the framework(FR) regions are those of a human immunoglobulin sequence. The humanizedantibody may optionally comprise at least a portion of a humanimmunoglobulin constant region (Fc).

The present invention includes anti-LAG3 fully human antibodies andantigen-binding fragments thereof and methods of use thereof. The term“fully human antibody” refers to an antibody that comprises humanimmunoglobulin protein sequences only. A fully human antibody maycontain murine carbohydrate chains if produced in a mouse, in a mousecell, or in a hybridoma derived from a mouse cell. Similarly, “mouseantibody” refers to an antibody that comprises mouse immunoglobulinsequences only. Alternatively, a fully human antibody may contain ratcarbohydrate chains if produced in a rat, in a rat cell, or in ahybridoma derived from a rat cell. Similarly, “rat antibody” refers toan antibody that comprises rat immunoglobulin sequences only. In anembodiment of the invention, an fully human anti-LAG3 antibody orantigen-binding fragment thereof is the product of isolation from atransgenic animal, e.g., a mouse (e.g., a HUMAB mouse, see e.g., U.S.Pat. Nos. 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016;5,770,429; 5,789,650; 5,814,318; 5,874,299 and 5,877,397; and Harding,et al., (1995) Ann. NY Acad. Sci. 764:536 546; or a XENOMOUSE, see e.g.,Green et al., 1999, J. Immunol. Methods 231:11-23), which has beengenetically modified to have fully human immunoglobulin genes; or theproduct of isolation from a phage or virus which expresses theimmunoglobulin chains of the anti-LAG3 fully human antibody orantigen-binding fragment thereof.

In general, the basic antibody structural unit comprises a tetramer.Each tetramer includes two identical pairs of polypeptide chains, eachpair having one “light” (about 25 kDa) and one “heavy” chain (about50-70 kDa). The amino-terminal portion of each chain includes a variableregion of about 100 to 110 or more amino acids primarily responsible forantigen recognition. The carboxy-terminal portion of the heavy chain maydefine a constant region primarily responsible for effector function.Typically, human light chains are classified as kappa and lambda lightchains. Furthermore, human heavy chains are typically classified as mu,delta, gamma, alpha, or epsilon, and define the antibody's isotype asIgM, IgD, IgG, IgA, and IgE, respectively. Within light and heavychains, the variable and constant regions are joined by a “J” region ofabout 12 or more amino acids, with the heavy chain also including a “D”region of about 10 more amino acids. See generally, FundamentalImmunology Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, N.Y. (1989).

In an embodiment of the invention, anti-LAG3 antibodies of the presentinvention comprise a full tetrameric structure having two light chainsand two heavy chains, including constant regions.

The variable regions of each light/heavy chain pair form the antibodybinding site. Thus, in general, an intact antibody has two bindingsites. Except in bifunctional or bispecific antibodies, the two bindingsites are, in general, the same.

Typically, the variable domains of both the heavy and light chainscomprise three hypervariable regions, also called complementaritydetermining regions (CDRs), located within relatively conservedframework regions (FR). The CDRs are usually aligned by the frameworkregions, enabling binding to a specific epitope. In general, fromN-terminal to C-terminal, both light and heavy chains variable domainscomprise FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The assignment ofamino acids to each domain is, generally, in accordance with thedefinitions of Sequences of Proteins of Immunological Interest, Kabat,et al.; National Institutes of Health, Bethesda, Md.; 5^(th) ed.; NIHPubl. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat,et al., (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al., (1987) JMol. Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883.

As used herein, the term “hypervariable region” refers to the amino acidresidues of an antibody or antigen-binding fragment thereof that areresponsible for antigen-binding. The hypervariable region comprisesamino acid residues from a “complementarity determining region” or “CDR”(i.e. CDR-L1, CDR-L2 and CDR-L3 in the light chain variable domain andCDR-H1, CDR-H2 and CDR-H3 in the heavy chain variable domain). See Kabatet al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed.Public Health Service, National Institutes of Health, Bethesda, Md.;Johnson et al. (2001) Nucleic Acids Res. 2001; 29(1): 205-206 (definingthe CDR regions of an antibody by sequence); see also Chothia and Lesk(1987) J. Mol. Biol. 196: 901-917; Chothia et al. Nature 342, 877(1989), and Tramontano et al. J. Mol. Biol. 215, 175 (1990) (definingthe CDR regions of an antibody by structure); see also Macallum et al. JMol Biol. 1996 Oct. 11; 262(5):732-45. As used herein, the term“framework” or “FR” residues refers to those variable domain residuesother than the hypervariable region residues defined herein as CDRresidues.

The scope of the present invention, includes anti-LAG3 antibodies andantigen-binding fragments thereof that specifically bind LAG3, whichhave any combination of CDRs from the immunoglobulin light chains of SEQID NOs: 7, 17, 27 and/or 37 and/or which have any combination of CDRsfrom the immunoglobulin heavy chains of SEQ ID NOs: 2, 12, 22 and 32wherein the CDRs are as defined by Kabat and Chothia (see above).

“Homology” refers to sequence similarity between two polynucleotidesequences or between two polypeptide sequences when they are optimallyaligned. When a position in both of the two compared sequences isoccupied by the same base or amino acid monomer subunit, e.g., if aposition in each of two DNA molecules is occupied by adenine, then themolecules are homologous at that position. The percent of homology isthe number of homologous positions shared by the two sequences dividedby the total number of positions compared ×100. For example, if 6 of 10of the positions in two sequences are matched or homologous when thesequences are optimally aligned then the two sequences are 60%homologous. Generally, the comparison is made when two sequences arealigned to give maximum percent homology.

“Isolated nucleic acid molecules” or “isolated polynucleotides” (e.g.,DNA or RNA) is also not associated with all or a portion of apolynucleotide in which the isolated polynucleotide is found in nature,or is linked to a polynucleotide to which it is not linked in nature.For purposes of this disclosure, it should be understood that “a nucleicacid molecule comprising” a particular nucleotide sequence does notencompass intact chromosomes. Isolated nucleic acid molecules“comprising” specified nucleic acid sequences may include, in additionto the specified sequences, coding sequences for up to ten or even up totwenty or more other proteins or portions or fragments thereof, or mayinclude operably linked regulatory sequences that control expression ofthe coding region of the recited nucleic acid sequences, and/or mayinclude vector sequences. As is discussed below, the present inventionincludes isolated polynucleotides encoding any of the immunoglobulinchains discussed herein.

The phrase “control sequences” refers to DNA sequences necessary for theexpression of an operably linked coding sequence in a particular hostorganism. The control sequences that are suitable for prokaryotes, forexample, include a promoter, optionally an operator sequence, and aribosome binding site. Eukaryotic cells are known to use promoters,polyadenylation signals, and enhancers.

A nucleic acid or polynucleotide is “operably linked” when it is placedinto a functional relationship with another nucleic acid sequence. Forexample, DNA for a presequence or secretory leader is operably linked toDNA for a polypeptide if it is expressed as a preprotein thatparticipates in the secretion of the polypeptide; a promoter or enhanceris operably linked to a coding sequence if it affects the transcriptionof the sequence; or a ribosome binding site is operably linked to acoding sequence if it is positioned so as to facilitate translation.Generally, but not always, “operably linked” means that the DNAsequences being linked are contiguous, and, in the case of a secretoryleader, contiguous and in reading phase. However, enhancers do not haveto be contiguous. Linking is accomplished by ligation at convenientrestriction sites. If such sites do not exist, the syntheticoligonucleotide adaptors or linkers are used in accordance withconventional practice.

As used herein, the expressions “cell,” and “cell line,” are usedinterchangeably and all such designations include progeny. Thus, thewords “transformants” and “transformed cells” include the primarysubject cell and cultures derived therefrom without regard for thenumber of transfers. It is also understood that not all progeny willhave precisely identical DNA content, due to deliberate or inadvertentmutations. Mutant progeny that have the same function or biologicalactivity as screened for in the originally transformed cell areincluded. Where distinct designations are intended, it will be clearfrom the context.

As used herein, “polymerase chain reaction” or “PCR” refers to aprocedure or technique in which specific nucleic acid sequences, RNAand/or DNA, are amplified as described in, e.g., U.S. Pat. No.4,683,195. Generally, sequence information from the ends of the regionof interest or beyond is used to design oligonucleotide primers. Theseprimers will be identical or similar in sequence to opposite strands ofthe template to be amplified. The 5′ terminal nucleotides of the twoprimers can coincide with the ends of the amplified material. PCR can beused to amplify specific RNA sequences, specific DNA sequences fromtotal genomic DNA, and cDNA transcribed from total cellular RNA,bacteriophage or plasmid sequences, etc. See generally Mullis et al.(1987) Cold Spring Harbor Symp. Quant. Biol. 51:263; Erlich, ed., (1989)PCR TECHNOLOGY (Stockton Press, N.Y.) As used herein, PCR is consideredto be one, but not the only, example of a nucleic acid polymerasereaction method for amplifying a nucleic acid test sample comprising theuse of a known nucleic acid as a primer and a nucleic acid polymerase toamplify or generate a specific piece of nucleic acid.

As used herein, “germline sequence” refers to a sequence of unrearrangedimmunoglobulin DNA sequences. Any suitable source of unrearrangedimmunoglobulin sequences may be used. Human germline sequences may beobtained, for example, from JOINSOLVER germline databases on the websitefor the National Institute of Arthritis and Musculoskeletal and SkinDiseases of the United States National Institutes of Health. Mousegermline sequences may be obtained, for example, as described inGiudicelli et al. (2005) Nucleic Acids Res. 33:D256-D261.

Physical and Functional Properties of the Exemplary Anti-LAG3 Antibodies

The present invention provides anti-LAG3 antibodies and antigen-bindingfragments thereof (e.g., humanized antibodies such as antagonisthumanized antibodies) and methods of use of the antibodies orantigen-binding fragments thereof in the treatment or prevention ofdisease. In one embodiment, the invention provides for mouse orhumanized anti-LAG3 antibodies and antigen-binding fragments thereof andmethods of use of the antibodies or antigen-binding fragments thereof inthe treatment or prevention of disease. In one embodiment, the inventionprovides for antagonistic anti-LAG3 antibodies and methods of use of theantibodies or antigen-binding fragments thereof in the treatment orprevention of disease.

Herein, an anti-LAG3 antibody or antigen-binding fragment thereofcomprising a particular light chain and a particular heavy chain may bereferred to as “light chain/heavy chain”; for example, an antibodycomprising the 45AGX_22D2_VL3 light chain and the Humanized×[LAG3_H]mAb.22D2 VH6 N54D heavy chain may be referred to as“45AGX_22D2_VL3/Humanized×[LAG3_H] mAb.22D2 VH6 N54D”.

An “anti-LAG3 antibody or antigen-binding fragment thereof” of thepresent invention includes any antibody or antigen-binding fragmentthereof that is discussed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) whichspecifically binds to LAG3 (e.g., human or cynomolgous monkey LAG3).Such antibodies and fragments include humanized antibodies and fragmentshaving any combination of the mouse or humanized light and heavy chainsthat are set forth herein or variants of such chains which specificallybind LAG3. Such antibodies and fragments include any antibody orfragment comprising any one or more of the CDRs (e.g., CDR-L1, CDR-L2,CDR-L3, CDR-H1, CDR-H2 and CDR-H3) of the mouse or humanized chains setforth herein or variants of such CDRs which specifically bind LAG3.Furthermore, an anti-LAG3 antibody or antigen-binding fragment thereofof the present invention includes any antibody or antigen-bindingfragment thereof that binds to the same epitope in LAG3 to which theantibodies and fragments discussed herein bind and any antibody orantigen-binding fragment that cross-blocks (partially or fully) or iscross-blocked (partially or fully) by an antibody or fragment discussedherein for LAG3 binding; as well as any variant thereof. A particularembodiment of the invention includes antibodies and fragments comprisingonly mouse immunoglobulin chains or only humanized immunoglobulin chainsand/or wherein the immunoglobulin chains or CDRs are all derived,directly or indirectly, from the same original mouse clone, i.e.,humanized 4A10 light chains paired with humanized 4A10 heavy chains;humanized 19E8 light chains paired with humanized 19E8 heavy chains;humanized 11C9 light chains paired with humanized 11C9 heavy chains; orhumanized 22D2 light chains paired with humanized 22D2 heavy chains; ormouse 4A10 light chains paired with mouse 4A10 heavy chains; mouse 19E8light chains paired with mouse 19E8 heavy chains; mouse 11C9 lightchains paired with mouse 11C9 heavy chains; or mouse 22D2 light chainspaired with mouse 22D2 heavy chains. These antibodies and fragments arepart of the present invention along with their uses, e.g., as set forthherein.

The cross-blocking antibodies and antigen-binding fragments thereofdiscussed herein can be identified based on their ability to block anyof the antibodies or fragments specifically set forth herein, e.g., Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, 4A10, 19E8, 11C9 and/or 22D2,from binding to LAG3, in binding assays (e.g., bio-layer interferometry(BLI; for example FORTEBIO OCTET binding assay; Pall ForteBio Corp;Menlo Park, Calif.), surface plasmon resonance (SPR), BIACore, ELISA,flow cytometry). For example, in an embodiment of the invention, whenusing BLI, the tip of a fiber-optic probe is coated with ligand (e.g.,LAG3) and acts as the biosensor wherein binding of anti-LAG3 antibody orantigen-binding fragment to the LAG3 alters the interference pattern ofwhite light reflected from the probe layer bound to LAG3 and an internalreference layer. The shift is indicative of LAG3/anti-LAG3 binding. Inan embodiment of the invention, the LAG3 coated tip is immersed in asolution of analyte containing antibody or antigen-binding fragment,e.g., in the well of either a 96- or 384-well plate. In an embodiment ofthe invention, the plate is shaken during reading to create orbitalflow. To read the assay, white light is directed down the length of thefiber. As mentioned above, interference between light reflecting off thereference layer and immobilized surfaces containing LAG3 of the tipcreates a distinctive pattern of light returning up the fiber. Asmolecules bind to the immobilized sensor surface, that pattern changesin proportion to the extent of binding. For example, assays can be usedin which a LAG3 (e.g., human LAG3) protein is immobilized on a BLI probeor plate, a reference anti-LAG3 antibody or fragment binds to LAG3(e.g., at saturating concentration) and a test anti-LAG3 antibody orfragment is added. The ability of the test antibody to compete with thereference antibody for LAG3 binding is then determined. In the BLIformat, light interference of the LAG3 complex is monitored to determineif the test antibody effectively competes with the reference antibody,e.g., nanometers of light wavelength shift over time is monitoredwherein a shift indicates additional binding of the test antibody and alack of cross-blocking. In an embodiment of the invention, in the BLIformat, cross-blocking is qualitatively deemed to have occurred betweenthe antibodies if no additional binding of test antibody is observed. Inan embodiment of the invention, as a control, cross-blocking of thereference antibody with itself is confirmed; wherein the assay isdetermined to be operating correctly if the reference antibody cancross-block itself from LAG3 binding. The ability of a test antibody toinhibit the binding of, for example, Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8, Ab9, 4A10, 19E8, 11C9 and/or 22D2, to LAG3 (e.g., human LAG3)demonstrates that the test antibody can cross-block Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8, Ab9, 4A10, 19E8, 11C9 and/or 22D2 for binding toLAG3 (e.g., human LAG3) and thus, may, in some cases, bind to the sameepitope on LAG3 (e.g., human LAG3) as Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8, Ab9, 4A10, 19E8, 11C9 and/or 22D2. As stated above, antibodies andfragments that bind to the same epitope as any of the anti-LAG3antibodies or fragments of the present invention also form part of thepresent invention. In an embodiment of the invention, BLI is conductedin a sandwich format wherein a reference anti-LAG3 antibody orantigen-binding fragment is immobilized to the probe and then bound withLAG3. Test anti-LAG3 antibody or antigen-binding fragment is then testedfor the ability to block binding of the references antibody or fragment.

“4A10”, “19E8”, “11C9” and “22D2” anti-LAG3 antibodies andantigen-binding fragments thereof referred to herein comprise theCDR-L1, CDR-L2 and CDR-L3 of the mouse immunoglobulin light chains 4A10,19E8, 11C9 or 22D2 and variants thereof, respectively; as well asCDR-H1, CDR-H2 and CDR-H3 of the mouse immunoglobulin heavy chains 4A10,19E8, 11C9 or 22D2, and variants thereof, respectively. Such “4A10”,“19E8”, “11C9” and “22D2” antibodies and fragments may be humanizedantibodies or antigen-binding fragments thereof such as Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9.

Examples of the immunoglobulin chains of anti-LAG3 antibodies as well astheir CDRs include, but are not limited to:

4A10-V_(H) sequenceATGAAATGCAGCTGGGTCATCTTCTTCCTGATGGCAGTGGTTATAGGAATCAATTCAGAGGTTCAGCTGCTCCAGTCTGGGGCAGAACTTGTGAGGTCAGGGGCCTCAGTCAAGTTGTCCTGCACAGCCTCTGGCTTCAACATTGAAGACTACTATATGCACTGGATGAAACAGAGGCCTGAACAGGGCCTGGAGTGGATTGGATGGATTGATCCTGTGAATGGTGATACTGAATATGCCCCGAAGTTCCAGGGCAAGGCCACTATGACTGCAGACACATCCTCCAACACAGCCTACCTACACCTCAACAGCCTGACATCTGAGGACACTGCCGTCTATTACTGTAATTTCTATGATGGTTACCTCTTTGCTTTCTGGGGCCAAGGGACCCTGGTCACTGTCTCTGCA (SEQ ID NO: 1; wherein the CDRs are underscored andwherein the signal sequence is in bold font)MKCSWVIFFLMAVVIGINSEVQLLQSGAELVRSGASVKLSCTASGFNIEDYY M HWMKQRPEQGLEWIGWIDPV NG DTEYAPKFQGKATMTADTSSNTAYLHLNSLTSEDTAVYYCNFYDGYLFAFWGQGTLVTVSA (SEQ ID NO: 2; whereinthe CDRs are underscored and wherein the signal sequence is in bold font)CDR-H1:  (SEQ ID NO: 3) GFNIEDYYMH CDR-H2:  (SEQ ID NO: 4)WIDPVNGDTEYAPKFQG CDR-H3:  (SEQ ID NO: 5) YDGYLFAF 4A10-V_(L) sequenceATGAGGTGCCTAGCTGAGTTCCTGGGGCTGCTTGTGCTCTGGATCCCTGGAGCCATTGGGGATATTGTGCTGACTCAGGCTGCACCCTCTGTACCTGTCACTCCTGGAGAGTCAGTGTCCATCTCCTGCAGGTCTAGTAAGAGTCTCCTGCATAGTGATGGCAACACTTATCTGTATTGGCTCCTGCAGAGGCCAGGCCAGTCTCCTCAGCTCCTGATATATCGGATGTCCAACCTTGCCTCAGGGGTCCCAGACAGGTTCAGCGGCAGTGGGTCAGGAACTGTTTTCACACTGAGAATCAGCAGACTGGAGGCTGAGGATGTGGGTATTTATTACTGTATGCAACATCTAGAATATCCTTTCACGTTTGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 6; wherein the CDRs are underscored and wherein thesignal sequence is in bold font)MRCLAEFLGLLVLWIPGAIGDIVLTQAAPSVPVTPGESVSISCRSSKSLLHSDGNTYLYWLLQRPGQSPQLLIYRM S NLASGVPDRFSGSGSGTVFTLRISRLEAEDVGIYYCMQHLEYPFTFGGGTKLEIK (SEQ ID NO: 7; whereinthe CDRs are underscored and wherein the signal sequence is in bold font)CDR-L1: (SEQ ID NO: 8) RSSKSLLHSDGNTYLY CDR-L2: (SEQ ID NO: 9) YRMSNLASCDR-L3: (SEQ ID NO: 10) MQHLEYPFT 19E8-V_(H) sequenceATGGGATGGAGCTGGATCTTTCTTTTCCTCCTGTCAGGAACTGCAGGTGTCCGTTGCCAGATCCGACTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGGTCCTCCTTCACTGACTACTATATAAACTGGGTGAAGCAGAAGCCTGGACAGGGACTTGAGTGGATTGGATGGATTTATCCTGGAAGCGGTAATTCTATCTACAATGAGAACTTCAAGGCCAAGGCCACATTGACTGTAGACACATCCTCCAGCACAGCCTACATGCATCTCAGCAGCCTGACATCTGAGGACACTGCTGTCTATTTCTGTGCAAGAGAGGCTGATTACGACGATGCTTTGGACTACTGGGGTCAAGGAACCTCGGTCACCGTCTCCTCA (SEQ ID NO: 11; wherein the CDRs are underscoredand wherein the signal sequence is in bold font)MGWSWIFLFLLSGTAGVRCQIRLQQSGPELVKPGASVKISCKASGSSFTDYYINWVKQKPGQGLEWIGWIYPGSG NSIYNENFKAKATLTVDTSSSTAYMHLSSLTSEDTAVYFCAREADYDDALDYWGQGTSVTVSS (SEQ ID NO: 12; wherein the CDRs are underscored and wherein the signal sequence is in bold font)CDR-H1: (SEQ ID NO: 13) GSSFTDYYIN CDR-H2: (SEQ ID NO: 14)WIYPGSGNSIYNENFKA CDR-H3: (SEQ ID NO: 15) EADYDDALDY 19E8-V_(L) sequenceATGGTATCCACACCTCAGTTCCTTGTATTTTTGCTTTTCTGGATTCCAGCCTCCAGAGGTCACATCTTGCTGACTCAGTCTCCAGCCATTCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGCATTGGCACAAGCATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATCCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCAGAAGATATTGCAGATTATTACTGTCAACAAAGTAATAGCTGGCCAACGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA(SEQ ID NO: 16; wherein the CDRs are underscored and wherein the signal sequenceis in bold font)MVSTPQFLVFLLFWIPASRGHILLTQSPAILSVSPGERVSFSCRASQSIGTSIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQS NS W PTYTEGGGTKLEIK(SEQ ID NO: 17; wherein the CDRs are underscored and wherein the signal sequenceis in bold font) CDR-L1: (SEQ ID NO: 18) RASQSIGTSIH CDR-L2:(SEQ ID NO: 19) YASESIS CDR-L3: (SEQ ID NO: 20) QQSNSWPTYT11C9-V_(H) sequenceATGAGATGGAGCTGTATCATCCTCTTCTTGGTAGCAACAGCTACAGGTGTCAACTCCCAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGATGCCTGGGGCTTCAGCGAAGATGTCCTGCAAGGCTTCTGGCTACACACTCACTGACTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATCGGAGCGATTGATATTTCTGATAGTTATTCTAGCTACAATCAAAAGTTCAAGGGCAAGGCCACATTGACTGTAGACGAATCCTCCAGCACAGCCTACATGCAGCTCACCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATCCCCTTTCTACAATAGTAGAGGGGGGAACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 21; wherein the CDRs areunderscored and wherein the signal sequence is in bold font)MRWSCIILFLVATATGVNSQVQLQQPGAELVMPGASAKMSCKASGYTLTDY W MHWVKQRPGQGLEWIGAIDISDSYSSYNQKFKGKATLTVDESSSTAYMQLTSLTSEDSAVYYCARSPFY NSRGGNYFDYWGQGTTLTVSS (SEQ ID NO: 22; wherein the CDRs are underscored and wherein thesignal sequence is in bold font) CDR-H1:  (SEQ ID NO: 23) GYTLTDYWMHCDR-H2: (SEQ ID NO: 24) AIDISDSYSSYNQKFKG CDR-H3: (SEQ ID NO: 25)SPFYNSRGGNYFDY 11C9-V_(L) sequenceATGATGTCCTCTGCTCAGTTCCTTGGTCTCCTGTTGCTCTGTTTTCAAGGTACCAGATGTGATATCCAGATGACACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACTACACATCAAGATTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAGAAGATATTGCCACTTACTTTTGCCAACAGGGTGATACGCTTCCTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA(SEQ ID NO: 26; wherein the CDRs are underscored and wherein the signal sequenceis in bold font)LAMSSAQFLGLLLLCFQGTRCDIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGDTLPPWTFGGGTKLEIK(SEQ ID NO: 27; wherein the CDRs are underscored and wherein the signal sequenceis in bold font) CDR-L1:  (SEQ ID NO: 28) RASQDISNYLN CDR-L2:(SEQ ID NO: 29) YTSRLHS CDR-L3: (SEQ ID NO: 30) QQGDTLPPWT22D2-V_(H) sequenceATGGGATGGACCTGGATCTTTCTCTTCTTCCTGTCAGGAACTGCAGGTGTCCTCTCTGAGGTCCTGCTGCTACAGTCTGGACCTGAACTGGTGAAGCCTGGGACTTCAGTGAAAATCCCCTGCAAGGCTTCTGGATACACATTCACTGACTACAACGTGGACTGGGTGAAGCAGCGCCATGGAAAGGGCCTTGAGTGGATTGGAGATATTAATCCAAACAATGGTGGTACTATCTACAGTCAGAAATTCAAGGGCAAGGCCACATTGACTGTTGACAAGTCCTCCAGCACAGCCTTCATGGAGCTCCGCAGCCTGACATCTGAGGACACTGCAGTCTATTTCTGTGCAAGGAACTATAGGTGGTTTGGTGCTATGGACCACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAACAGCCCCATCGGTCTATCCACTG(SEQ ID NO: 31; wherein the CDRs are underscored and wherein the signal sequenceis in bold font)MGWTWIFLFFLSGTAGVLSEVLLLQSGPELVKPGTSVKIPCKASGYTFTDYNVDWVKQRHGKGLEWIGDI NP N NG GTIYSQKFKGKATLTVDKSSSTAFMELRSLTSEDTAVYFCARNYR W FGAMDHWGQGTSVTVSS(SEQ ID NO: 32; wherein the CDRs are underscored and wherein the signal sequenceis in bold font) CDR-H1: (SEQ ID NO: 33) DYNVD CDR-H2: (SEQ ID NO: 34)DINPNNGGTIYSQKFKG CDR-H3: (SEQ ID NO: 35) NYRWFGAMDH 22D2-V_(L) sequenceATGGAGACAGACACAATCCTGCTATGGGTGCTGCTGCTCTGGGTTCCAGGTTCCACTGGTGACATTGTGTTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCCAGGGCAGAGGGCCACCATTTCCTGCAAGGCCAGTCAAAGTCTTGATTATGAAGGTGATAGTGATATGAATTGGTACCAACAGAAACCAGGACAGCCACCCAGACTCCTCATCTCTGGTGCATCCAATCTAGAGTCTGGGATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTGTTAACATCCATCCTGTGGAGGAGGAGGATGCTGCAACCTATTACTGTCAGCAAAGTACTGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGGGCTGATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCAGTTAACATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAAAGACATCAATGTCAAGTGGAAGATTGATGGCAGTGAACGACAAAATGGCG (SEQ ID NO: 36; wherein the CDRs are underscored and wherein thesignal sequence is in bold font)METDTILLWVLLLWVPGSTGDIVLTQSPASLAVSPGQRATISCKASQSLDYEGDSDMNWYQQKPGQPPRLLISGASNLESGIPARFSGSGSGTDFTVNIHPVEEEDAATYYCQQSTEDPRTFGGGTKLEIK (SEQ ID NO: 37; whereinthe CDRs are underscored and wherein the signal sequence is in bold font)CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN CDR-L2: (SEQ ID NO: 39) GASNLESCDR-L3: (SEQ ID NO: 40) QQSTEDPRT

The 22D2 mouse parental heavy or light chains may be referred to hereinas LB145.22D2.E1.1D1. The 19E8 mouse parental heavy or light chains maybe referred to herein as LB148.19E8.G1.1A1. The 4A10 mouse parentalheavy or light chains may be referred to herein as LB148.4A10.1H1. The11C9 mouse parental heavy or light chains may be referred to herein asLB148.11C9.1C1.

The present invention also includes any anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibodies such asantagonist humanized antibodies) comprising one or more of the heavyand/or light chains (or variants thereof) or CDRs (or variants thereof)or mature fragments of such chains (or variants thereof) or variabledomains thereof of such chains (or variants thereof) which are set forthe below. Light chains may be designated with a “V_(L)” or “VK” andheavy chains may be designated with a “V_(H)”.

Mouse Immunoglobulin Chains

Chains set forth below having a “4A10”, “19E8”, “11C9” or “22D2”designation may be referred to as such herein. As discussed herein, thescope of the present invention includes anti-LAG3 antibodies (e.g.,humanized antibodies such as humanized antagonistic antibodies) andantigen-binding fragments thereof comprising any one or more (e.g., 3)light chain CDRs and/or any one or more (e.g., 3) heavy chain CDRs fromthe immunoglobulin chains set forth below; or any mature variable domainof a light immunoglobulin chain and/or mature variable domain of a heavyimmunoglobulin chain set forth in SEQ ID NOs: 45-104.

Humanized Chains

In an embodiment of the invention, a humanized (e.g., humanizedantagonistic) anti-LAG3 antibody or antigen-binding fragment of theinvention comprises any combination of heavy and light mature, variabledomains of the following immunoglobulin chains. In an embodiment of theinvention, the 11C9 humanized light chains are paired with the 11C9humanized heavy chains; the 19E8 humanized light chains are paired withthe 19E8 humanized heavy chains; the 4A10 humanized light chains arepaired with the 4A10 humanized heavy chains; and the 22D2 humanizedlight chains are paired with the 22D2 humanized heavy chains. In anembodiment of the invention, a humanized anti-LAG3 antibody orantigen-binding fragment thereof comprises a “45AGX_22D2_VL3”immunoglobulin variable domain light chain, e.g., comprising the aminoacid sequence of SEQ ID NO: 274 or a mature fragment thereof (e.g.,amino acids 21-131 of SEQ ID NO: 274); and a “Humanized×[LAG3_H]mAb.22D2 VH6 N54D” or “Humanized×[LAG3_H] mAb.22D2 VH6 N54G”immunoglobulin variable domain heavy chain comprising the amino acidsequence of SEQ ID NO: 426 or SEQ ID NO: 427, respectively.

Chains set forth below having a “4A10”, “19E8”, “11C9” or “22D2”designation may be referred to as such herein. As discussed herein, thescope of the present invention includes anti-LAG3 antibodies (e.g.,humanized antibodies such as humanized antagonistic antibodies) andantigen-binding fragments thereof comprising any one or more (e.g., 3)light chain CDRs and/or any one or more (e.g., 3) heavy chain CDRs fromthe immunoglobulin chains set forth herein; or any mature or unprocessedV_(L) domain or light chain immunoglobulin and/or mature or unprocessedV_(H) domain or heavy chain immunoglobulin set forth herein. The scopeof the present invention also includes any of the humanized polypeptidesor polynucleotides or variable domains thereof having 1, 2, 3, 4, 5, 6,7, 8, 9 or 10 point mutations or point deletions.

Specific embodiments of the invention include any humanized anti-LAG3antibodies and antigen-binding fragments thereof comprising theimmunoglobulin light and heavy chains set forth below or any antibody orfragment having the light and heavy chain CDRs thereof (e.g., IgG1 orIgG4). Such antibodies and fragments may be referred to as any one ofAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9 as follows:

-   -   Ab1: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 53AHH Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6) IgG1/Kappa (PX) (or the variable domain        thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 106) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 106 (CDRs underscored))or comprising the CDRs:

CDR-L1:  (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2:  (SEQ ID NO: 39)GASNLES; CDR-L3:  (SEQ ID NO: 40) QQSTEDPRT; CDR-H1:  (SEQ ID NO: 33)DYNVD; CDR-H2:  (SEQ ID NO: 458) DINPNNGGTIYAQKFQE; and CDR-H3: (SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab2: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 56AHH Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55S) IgG1/Kappa (PX) (or the variable        domain thereof); for example: comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (amino acids 21-238 of SEQ ID NO: 126);and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 108) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 108 (CDRs underscored))or comprising the CDRs:

CDR-L1:  (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2:  (SEQ ID NO: 39)GASNLES; CDR-L3:  (SEQ ID NO: 40) QQSTEDPRT; CDR-H1:  (SEQ ID NO: 33)DYNVD; CDR-H2:  (SEQ ID NO: 456) DINPNSGGTIYAQKFQE; and CDR-H3: (SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab3: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 54AHH Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55D) IgG1/Kappa (PX) (or the variable        domain thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC;

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 110) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 110 (CDRs underscored));or comprising the CDRs:

CDR-L1:  (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2:  (SEQ ID NO: 39)GASNLES; CDR-L3:  (SEQ ID NO: 40) QQSTEDPRT; CDR-H1:  (SEQ ID NO: 33)DYNVD; CDR-H2:  (SEQ ID NO: 457) DINPNDGGTIYAQKFQE; and CDR-H3: (SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab4: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 52AHH Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55Q) IgG1/Kappa (PX) (or the variable        domain thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 112) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 112 (CDRs underscored));or comprising the CDRs:

CDR-L1:  (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2:  (SEQ ID NO: 39)GASNLES; CDR-L3:  (SEQ ID NO: 40) QQSTEDPRT; CDR-H1:  (SEQ ID NO: 33)DYNVD; CDR-H2:  (SEQ ID NO: 455) DINPNQGGTIYAQKFQE; and CDR-H3: (SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab5: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 57AHH Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6) IgG4 S228P (PX) (or the variable domain        thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC;and (SEQ ID NO: 114) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK,or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNNGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 114 (CDRs underscored));or comprising the CDRs:

CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2: (SEQ ID NO: 39)GASNLES; CDR-L3: (SEQ ID NO: 40) QQSTEDPRT; CDR-H1: (SEQ ID NO: 33)DYNVD; CDR-H2: (SEQ ID NO: 458) DINPNNGGTIYAQKFQE; and CDR-H3:(SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab6: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 73AHD Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55D/VL3) IgG4 S228P/Kappa (PX) (or the        variable domain thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 116) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored))DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQST EDPRTFGGGTKVEIK;and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 1-119 of SEQ ID NO: 116 (CDRs underscored))QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNDGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS;or comprising the CDRs:

CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2: (SEQ ID NO: 39)GASNLES; CDR-L3: (SEQ ID NO: 40) QQSTEDPRT; CDR-H1: (SEQ ID NO: 33)DYNVD; CDR-H2: (SEQ ID NO: 457) DINPNDGGTIYAQKFQE; and CDR-H3:(SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab7: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 21AHG Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55S/VL3) IgG4 S228P/Kappa (PX) (or the        variable domain thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 118) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored))DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQST EDPRTFGGGTKVEIK;and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 1-119 of SEQ ID NO: 118 (CDRs underscored))QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNSGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS;or comprising the CDRs:

CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2: (SEQ ID NO: 39)GASNLES; CDR-L3: (SEQ ID NO: 40) QQSTEDPRT; CDR-H1: (SEQ ID NO: 33)DYNVD; CDR-H2: (SEQ ID NO: 456) DINPNSGGTIYAQKFQE; and CDR-H3:(SEQ ID NO: 35) NYRWFGAMDH

-   -   Ab8: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 80AHG Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55Q/VL3) IgG4 S228P/Kappa (PX) (or the        variable domain thereof); for example comprising:

-   a light chain immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 120) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALH NHYTQKSLSLSLGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored))DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQST EDPRTFGGGTKVEIK;and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

(amino acids 1-119 of SEQ ID NO: 120 (CDRs underscored))QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNQGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS;or comprising the CDRs:

CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2: (SEQ ID NO: 39)GASNLES; CDR-L3: (SEQ ID NO: 40) QQSTEDPRT; CDR-H1: (SEQ ID NO: 33)DYNVD; CDR-H2: (SEQ ID NO: 455) DINPNQGGTIYAQKFQE; and CDR-H3:(SEQ ID NO: 35) NYRWFGAMDHor

-   -   Ab9: humanized light chain 45AGX Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 (VL3)) Kappa (PX) (or the variable domain        thereof) and humanized heavy chain 72AHD Humanized×[LAG3_H] mAb        (LB145.22D2.E1.D1 VH6 N55G/VL3) IgG4 S228P/Kappa (PX)) (or the        variable domain thereof); for example comprising: a light chain        immunoglobulin comprising the amino acid sequence:

(amino acids 21-238 of SEQ ID NO: 126)DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC;and

-   a heavy chain immunoglobulin comprising the amino acid sequence:

(SEQ ID NO: 122) QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNGGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK;or

-   a light chain immunoglobulin variable domain comprising the amino    acid sequence:

DIVMTQTPLSLSVTPGQPASISCKASQSLDYEGDSDMNWYLQKPGQPPQLLIYGASNLESGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSTEDPRTFGGGTKVEIK (amino acids 21-131 of SEQ ID NO: 126 (CDRs underscored));and

-   a heavy chain immunoglobulin variable domain comprising the amino    acid sequence:

QMQLVQSGPEVKKPGTSVKVSCKASGYTFTDYNVDWVRQARGQRLEWIGDINPNGGGTIYAQKFQERVTITVDKSTSTAYMELSSLRSEDTAVYYCARNYRWFGAMDHWGQGTTVTVSS (amino acids 1-119 of SEQ IDNO: 122 (CDRs underscored));or comprising the CDRs:

CDR-L1: (SEQ ID NO: 38) KASQSLDYEGDSDMN; CDR-L2: (SEQ ID NO: 39)GASNLES; CDR-L3: (SEQ ID NO: 40) QQSTEDPRT; CDR-H1: (SEQ ID NO: 33)DYNVD; CDR-H2: (SEQ ID NO: 455) DINPNQGGTIYAQKFQE; and CDR-H3:(SEQ ID NO: 35) NYRWFGAMDH

In an embodiment of the invention, the CDR-H2 of any anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention comprisesthe amino acid sequence:

(SEQ ID NO: 446) DINPNX₁GGTIYX₂QKFX₃X₄wherein,

-   X₁=D, N, S or Q-   X₂=A or S-   X₃=Q or K-   X₄=E or G

Humanized heavy immunoglobulin chains are set forth in SEQ ID NOs: 106,108, 110, 112, 114, 116, 118, 120, 122, 124, 124, 128, 134, 140, 142,144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170,172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198,200, 202, 204, 206, 212, 214, 216, 218, 220, 222, 234, 235, 237, 239,243, 245, 247, 249, 251, 253, 255, 265, 267, 269, 271, 273, 275, 277,279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333,335, 337, 339, 341, 343, 345, 347, 349, 353, 355, 357, 359, 361, 363,365, 367, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399,406-419, 448, 449, 462 and 463. DNA encoding humanized heavyimmunoglobulin chains are set forth in SEQ ID NOs: 105, 107, 109, 111,113, 115, 117, 119, 121, 123, 127, 133, 139, 141, 143, 145, 147, 149,151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177,179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205,211, 213, 215, 217, 219, 221, 233, 236, 238, 242, 244, 246, 248, 250,252, 254, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286,288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314,316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342,344, 346, 348, 352, 354, 356, 358, 360, 362, 364, 366, 376, 378, 380,382, 384, 386, 388, 390, 392, 394, 396 and 398.

Humanized light immunoglobulin chains are set forth in SEQ ID NOs: 126,130, 132, 136, 138, 208, 210, 224, 226, 228, 230, 232, 241, 257, 259,261, 263, 351, 369, 371, 373, 375, 401, 403, 405, 450-453, 426, 427 and459-461. DNA encoding humanized light immunoglobulin chains are setforth in SEQ ID NOs: 125, 129, 131, 135, 137, 207, 209, 223, 225, 227,229, 231, 240, 256, 258, 260, 262, 350, 368, 370, 372, 374, 400, 402 and404.

A “variant” of a polypeptide, such as an immunoglobulin chain, refers toa polypeptide comprising an amino acid sequence that is at least about70-99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87,88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) identicalor similar to a referenced amino acid sequence that is set forth herein;when the comparison is performed by a BLAST algorithm wherein theparameters of the algorithm are selected to give the largest matchbetween the respective sequences over the entire length of therespective reference sequences (e.g., expect threshold: 10; word size:3; max matches in a query range: 0; BLOSUM 62 matrix; gap costs:existence 11, extension 1; conditional compositional score matrixadjustment).

A “variant” of a polynucleotide refers to a polynucleotide comprising anucleotide sequence that is at least about 70-99.9% (e.g., 70, 72, 74,75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 99.5, 99.9%) identical to a referenced nucleotidesequence that is set forth herein; when the comparison is performed by aBLAST algorithm wherein the parameters of the algorithm are selected togive the largest match between the respective sequences over the entirelength of the respective reference sequences (e.g., expect threshold:10; word size: 28; max matches in a query range: 0; match/mismatchscores: 1, −2; gap costs: linear).

Polypeptides and anti-LAG3 antibodies and antigen-binding fragmentsthereof (e.g., humanized antibodies) of the present invention, in anembodiment of the invention, include a heavy chain immunoglobulinvariable region having at least 78.99% (e.g., 79%, 80%, 85%, 90%, 95%,99%) amino acid sequence identity to amino acids 1-119 of SEQ ID NO:106; and/or a light chain immunoglobulin variable region having at least78.38% (e.g., 79%, 80%, 85%, 90%, 95%, 99%) amino acid sequence identityto amino acids 1-111 of SEQ ID NO: 224.

In addition, a variant may be a polypeptide comprising an amino acidsequence that is set forth herein except for one or more (e.g., 1, 2, 3,4, 5, 6, 7, 8, 9 or 10) mutations such as, for example, missensemutations (e.g., conservative substitutions), non-sense mutations,deletions, or insertions. Such a polypeptide may be an immunoglobulinlight chain, an immunoglobulin heavy chain and/or a CDR (e.g., any oneor more of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and/or CDR-H3).

As discussed herein, the present invention includes anti-LAG3 antibodiesand antigen-binding fragments thereof that include one or more variantsof the framework sequences (e.g., any one or more of FR-L1, FR-L2,FR-L3, FR-L4, FR-H1, FR-H2, FR-H3 and/or FR-H4), CDRs (e.g., 1, 2 or 3variant CDR-Ls and/or 1, 2, or 3 variant CDR-Hs) and/or immunoglobulinchains (e.g., 1 or 2 variant V_(L)s and/or 1 or 2 variant V_(H)s) whosesequences are specifically set forth herein. Such antibodies andantigen-binding fragments may, themselves, be referred to as variants.Simple polypeptide chains, that include one or more variant FRs, CDR-Ls,CDR-Hs and/or immunoglobulin chains, themselves are also part of thepresent invention. Polynucleotides encoding such variant polypeptidechains are also part of the present invention. For example, the presentinvention provides anti-LAG3 antibodies and antigen-binding fragmentsthereof that comprising the amino acid sequence of the V_(H) and V_(L)of the antibodies Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9, whichare set forth herein, as well as variants thereof comprising the CDR-L1,CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of said Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 or Ab9 but comprising 70% or more (e.g., 80%, 85%,90%, 95%, 97% or 99%) overall amino acid sequence identity or similarityto said V_(H) and V_(L) of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 orAb9. Thus, in such embodiments, the CDRs of the antibodies and fragmentsare identical to those of the V_(H) and V_(L) of Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 or Ab9 but any differences from such V_(H) and V_(L)occur in the frameworks and/or immunoglobulin constant domains.

The invention provides variant anti-LAG3 antibodies or antigen-bindingfragments thereof (e.g., humanized antibodies such as antagonisthumanized antibodies) comprising one or more variant CDRs (e.g., any oneor more of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and/or CDR-H3) and/orframework regions (e.g., any one or more of FR1, FR2, FR3 and/or FR4)that are set forth herein; and/or one or more variant V_(L) domainsand/or one or more variant V_(H) domains of such antibodies or fragmentsthat are set forth herein, for example, with at least 70%, 75%, 80%,85%, 90%, 95%, 98%, 99% or 99.9% sequence identity or similarity to,e.g., SEQ ID NO: 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14, 15, 17, 18, 19,20, 22, 23, 24, 25, 27, 28, 29, 30, 32, 33, 34, 35, 37, 38, 39, 40, 45,47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81,83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 106, 108, 110, 112, 114,116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142,144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170,172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198,200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226,228, 230, 232, 234, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253,255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281,283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309,311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337,339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365,367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393,395, 397, 399, 401, 403, 405, 406, 407, 408, 409, 410, 411, 412, 413,414, 415, 416, 417, 418, 419, 426, 427, 434, 435, 436, 437, 438, 439,440, 441, 442, 446, 448, 449, 451, 452, 453, 454, 455, 456, 457, 458,459, 460, 461, 462, 463 or 464; which specifically bind to LAG3.

As discussed above, the scope of the present invention includes variantanti-LAG3 antibodies or antigen-binding fragments comprising one or morevariant CDRs (e.g., 1, 2 or 3 variant CDR-Ls and/or 1, 2, or 3 variantCDR-H); and/or framework regions (e.g., any one or more of FR1, FR2, FR3and/or FR4) and/or variant V_(L) and/or variant V_(H) domains (with orwithout a signal sequence) having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or moremutations. The mutations can include point mutations which areconservative or non-conservative amino acid substitutions or pointdeletions, for example in a framework region and/or in a CDR. Asdiscussed above, the present invention provides anti-LAG3 antibodies andantigen binding fragments thereof comprising CDR-L1, CDR-L2, CDR-L3,CDR-H1, CDR-H2 and CDR-H3 of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 orAb9 but having mutations in the framework regions thereof.

Conservatively modified variant anti-LAG3 antibodies and antigen-bindingfragments thereof are also part of the present invention. A“conservatively modified variant” or a “conservative substitution”refers to a variant wherein there is one or more substitutions of aminoacids in a polypeptide with other amino acids having similarcharacteristics (e.g. charge, side-chain size,hydrophobicity/hydrophilicity, backbone conformation and rigidity,etc.). Such changes can frequently be made without significantlydisrupting the biological activity of the antibody or fragment. Those ofskill in this art recognize that, in general, single amino acidsubstitutions in non-essential regions of a polypeptide do notsubstantially alter biological activity (see, e.g., Watson et al. (1987)Molecular Biology of the Gene, The Benjamin/Cummings Pub. Co., p. 224(4th Ed.)). In addition, substitutions of structurally or functionallysimilar amino acids are less likely to significantly disrupt biologicalactivity.

Variant anti-LAG3 antibodies or antigen-binding fragments of the presentinvention, which are discussed herein, comprise one or more CDRs (e.g.,1, 2 or 3 variant CDR-Ls and/or 1, 2, or 3 variant CDR-H); frameworkregions (e.g., any one or more of FR1, FR2, FR3 and/or FR4); and/orimmunoglobulin chains having one or more conservative substitutions. Forexample, such antibodies and fragments may comprise the amino acidsequences disclosed herein, e.g. SEQ ID NOs: 2, 3, 4, 5, 7, 8, 9, 10,12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 27, 28, 29, 30, 32, 33,34, 35, 37, 38, 39, 40, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67,69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101,103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130,132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158,160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186,188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214,216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 237, 239, 241,243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269,271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297,299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325,327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353,355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381,383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 406, 407,408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 426, 427,434, 435, 436, 437, 438, 439, 440, 441, 442, 446, 448, 449, 451, 452,453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463 or 464; whereinsuch amino acid sequences may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,12, 15, 20 or more conservative amino acid substitutions thereof.Exemplary conservative substitutions are set forth in Table 1.

TABLE 1 Exemplary Conservative Amino Acid Substitutions Original residueConservative substitution Ala (A) Gly; Ser Arg (R) Lys; His Asn (N) Gln;His Asp (D) Glu; Asn Cys (C) Ser; Ala Gln (Q) Asn Glu (E) Asp; Gln Gly(G) Ala His (H) Asn; Gln Ile (I) Leu; Val Leu (L) Ile; Val Lys (K) Arg;His Met (M) Leu; Ile; Tyr Phe (F) Tyr; Met; Leu Pro (P) Ala Ser (S) ThrThr (T) Ser Trp (W) Tyr; Phe Tyr (Y) Trp; Phe Val (V) Ile; Leu

Function-conservative variants of the anti-LAG3 antibodies andantigen-binding fragments thereof are also part of the presentinvention. Any of the variants of the anti-LAG3 antibodies andantigen-binding fragments thereof (as discussed herein) may be“function-conservative variants”. Such function-conservative variantsmay, in some cases, also be characterized as conservatively modifiedvariants. “Function-conservative variants,” as used herein, refers tovariants of the anti-LAG3 antibodies or antigen-binding fragmentsthereof in which one or more amino acid residues (e.g., of 1, 2, 3, 4, 5or 6 CDRs and/or of a V_(L) and/or of a V_(H)) have been changed withoutsignificantly altering one or more functional properties of the antibodyor fragment. In an embodiment of the invention, a function-conservativevariant anti-LAG3 antibody and antigen-binding fragments thereof of thepresent invention (e.g., humanized antibodies such as antagonisthumanized antibodies) comprise a variant of an immunoglobulin chain(e.g., one or two variant V_(H)s and/or one or more variant V_(L)s)and/or of a CDR (e.g., 1, 2 or 3 variant CDR-Ls and/or 1, 2, or 3variant CDR-H) of any of those set forth herein, e.g., any of SEQ IDNOs: 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23,24, 25, 27, 28, 29, 30, 32, 33, 34, 35, 37, 38, 39, 40, 45, 47, 49, 51,53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87,89, 91, 93, 95, 97, 99, 101, 103, 106, 108, 110, 112, 114, 116, 118,120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146,148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174,176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202,204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230,232, 234, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257,259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285,287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313,315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341,343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369,371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397,399, 401, 403, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415,416, 417, 418, 419, 426, 427, 434, 435, 436, 437, 438, 439, 440, 441,442, 446, 448, 449, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460,461, 462, 463 or 464; exhibiting one or more of the following functionalproperties:

-   -   Inhibits LAG3 binding to MHC class II molecules, e.g., on Daudi        cells; for example inhibits human LAG3/MHC class II binding on        Daudi cells with an IC₅₀ of about 1.9-2.9 nM, e.g., 2.1 nM, 2.8        nM, 2.0 nM, 1.9 nM, 2.5 nM, 2.6 nM, 2.1 nM, 2.4 nM or 2.5 nM.        (e.g., about 2.1-2.6 nM).    -   Competes with MEW class II molecules for LAG3 binding e.g., on        Daudi cells;    -   Binds the extraloop of LAG3;    -   Binds LAG3 with a K_(D) of about 10⁻⁹M to about 2×10⁻¹²M        affinity (e.g., as measured by surface plasmon resonance or        KinExA); for example binds human LAG3 with a KD of about 2, 3,        6, 10 or 11 pM (e.g., 2-11 pM) and/or binds cynomolgous monkey        LAG3 with a KD of about 11, 12, 16 or 25 pM (e.g., 11-25 pM),        e.g., by KinExA;    -   Binds to native LAG3 on the surface of activated CD4+ and/or        CD8+ T-cells; for example, binds human CD4+ T-cells expressing        human LAG3 with an EC₅₀ of about 39, 41 or 57 pM (e.g., about        39-57 pM); binds human CD8+ T-cells expressing human LAG3 with        an EC₅₀ of about 33, 35 or 49 pM (e.g., about 33-49 pM); binds        cynomolgous monkey CD4+ T-cells expressing cynomolgous monkey        LAG3 with an EC₅₀ of about 27, 30 or 35 pM (e.g., about 27-35        pM); binds cynomolgous monkey CD8+ T-cells expressing        cynomolgous monkey LAG3 with an EC₅₀ of about 30, 31 or 41 pM        (e.g., about 30-41 pM); for example wherein the T-cells are        isolated from blood;    -   Binds to human and/or cynomolgous monkey, e.g., Macaca        fascicularis or Macaca mulatta LAG3;    -   Inhibits LAG3 homodimerization;    -   Stimulates antigen-specific T-cell production of IL-2, e.g., IL2        production from the 3A9 murine T-cell hybridoma expressing human        LAG3 with an EC₅₀ of about 1.06-1.65 nM, 1.74-1.83 nM, 3.56-4.06        nM, 2.83-2.96 nM, 0.57-1.07 nM, 0.45-1.27 nM, 0.47-1.01 nM or        0.72-1.08 nM;    -   labels tonsil tissue; and/or    -   enhances T-cell activation by anti-PD1 antibody such as        pembrolizumab, e.g., increases IL-2 production by T-cells;    -   does not label brain, heart, kidney, liver, lung, pancreas,        and/or pituitary tissue.    -   binds to human LAG3 by contacting residues QEGAPAQL (amino acids        35-42 of SEQ ID NO: 443) and RPARRADAGEYRAAVH (amino acids        137-152 of SEQ ID NO: 443) and, optionally, residues        DERGRQRGDFSLW (amino acids 123-135 of SEQ ID NO: 443) of LAG3;        or residues SPTIPLQDL (amino acids 45-53 of SEQ ID NO: 443) and,        optionally DERGRQRGDFSL (amino acids 123-134 of SEQ ID NO: 443)        of LAG3; or residues HPLAPGPHPAAPSSWGPRPRRYTVL (amino acids        78-102 of SEQ ID NO: 443) of LAG3; and/or by protecting        hydrogens on the amide backbone of such residues from exchange        with a deuterium (e.g., from D₂O).

The present invention provides a method for making an antibody orantigen-binding fragment thereof that binds specifically to LAG3comprising administering, to a non-human host animal (e.g., mouse,rabbit, camel, llama or rat), an effective amount of one or morepeptides comprising, consisting of or consisting essentially of an aminoacid sequence selected from the group consisting of QEGAPAQL (aminoacids 35-42 of SEQ ID NO: 443); RPARRADAGEYRAAVH (amino acids 137-152 ofSEQ ID NO: 443); DERGRQRGDFSLW (amino acids 123-135 of SEQ ID NO: 443);SPTIPLQDL (amino acids 45-53 of SEQ ID NO: 443); DERGRQRGDFSL (aminoacids 123-134 of SEQ ID NO: 443); and HPLAPGPHPAAPSSWGPRPRRYTVL (aminoacids 78-102 of SEQ ID NO: 443), e.g., wherein the peptide is formulatedwith a pharmaceutically acceptable carrier. Optionally, the antibody orfragment is isolated from the host animal, e.g., from the serum or bloodof the host animal. Optionally, the host animal is administered morethan one dose of the peptide. Such isolated peptides are part of thepresent invention, e.g., fused to an immunogen such as Keyhole LimpetHemocyanin (KLH), human serum albumin or bovine serum albumin.

Sequence identity refers to the degree to which the amino acids of twopolypeptides are the same at equivalent positions when the two sequencesare optimally aligned. Sequence similarity includes identical residuesand nonidentical, biochemically related amino acids. Biochemicallyrelated amino acids that share similar properties and may beinterchangeable are discussed above.

The following references relate to BLAST algorithms often used forsequence analysis: BLAST ALGORITHMS: Altschul et al. (2005) FEBS J.272(20): 5101-5109; Altschul, S. F., et al., (1990) J. Mol. Biol.215:403-410; Gish, W., et al., (1993) Nature Genet. 3:266-272; Madden,T. L., et al., (1996) Meth. Enzymol. 266:131-141; Altschul, S. F., etal., (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J., et al., (1997)Genome Res. 7:649-656; Wootton, J. C., et al., (1993) Comput. Chem.17:149-163; Hancock, J. M. et al., (1994) Comput. Appl. Biosci.10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, M. O., et al., “A model ofevolutionary change in proteins.” in Atlas of Protein Sequence andStructure, (1978) vol. 5, suppl. 3. M. O. Dayhoff (ed.), pp. 345-352,Natl. Biomed. Res. Found., Washington, D.C.; Schwartz, R. M., et al.,“Matrices for detecting distant relationships.” in Atlas of ProteinSequence and Structure, (1978) vol. 5, suppl. 3.” M. O. Dayhoff (ed.),pp. 353-358, Natl. Biomed. Res. Found., Washington, D.C.; Altschul, S.F., (1991) J. Mol. Biol. 219:555-565; States, D. J., et al., (1991)Methods 3:66-70; Henikoff, S., et al., (1992) Proc. Natl. Acad. Sci. USA89:10915-10919; Altschul, S. F., et al., (1993) J. Mol. Evol.36:290-300; ALIGNMENT STATISTICS: Karlin, S., et al., (1990) Proc. Natl.Acad. Sci. USA 87:2264-2268; Karlin, S., et al., (1993) Proc. Natl.Acad. Sci. USA 90:5873-5877; Dembo, A., et al., (1994) Ann. Prob.22:2022-2039; and Altschul, S. F. “Evaluating the statisticalsignificance of multiple distinct local alignments.” in Theoretical andComputational Methods in Genome Research (S. Suhai, ed.), (1997) pp.1-14, Plenum, N.Y.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention (e.g., humanized antibodies such as antagonisthumanized antibodies) can comprise one, two, three, four, five, or sixof the complementarity determining regions (CDRs) of the immunoglobulinchains disclosed herein (wherein 1, 2, 3, 4, 5 or 6 of the CDRs are,optionally, variants of those set forth herein). The one, two, three,four, five, or six CDRs may be independently selected from the CDRsequences of the various immunoglobulin chains disclosed herein.Alternatively, the one, two, three, four, five, or six CDRs may beselected from the CDR sequences of a single described antibody of theinvention.

For example, the present invention includes anti-LAG3 antibodies andantigen-binding fragments thereof as well as immunoglobulin polypeptidechains that comprise:

-   -   the 4A10 CDR-H1, CDR-H2 and CDR-H3;    -   the 4A10 CDR-L1, CDR-L2 and CDR-L3;    -   the 11C9 CDR-H1, CDR-H2 and CDR-H3;    -   the 11C9 CDR-L1, CDR-L2 and CDR-L3;    -   the 19E8 CDR-H1, CDR-H2 and CDR-H3;    -   the 19E8 CDR-L1, CDR-L2 and CDR-L3;    -   the 22D2 CDR-H1, CDR-H2 and CDR-H3; and/or    -   the 22D2 CDR-L1, CDR-L2 and CDR-L3; wherein the 4A10, 11C9, 19E8        and 22D2 CDRs may be derived from the mouse or humanized 4A10,        11C9, 19E8 and 22D2 immunoglobulin chains, respectively, set        forth herein (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or        Ab9).

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody heavy chainvariable (V_(H)) domain comprising one or more (e.g., 3) of CDR-H1,CDR-H2 or CDR-H3 of 4A10 V_(H) (e.g., SEQ ID NO: 2); e.g., wherein theCDRs comprise the amino acid sequences set forth in SEQ ID NOs: 3 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), 4 (or a variant thereof having 1, 2, 3, 4, 5,6, 7, 8, 9 or 10 point mutations and/or point deletions), and 5 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody light chainvariable (V_(L)) domain comprising one or more (e.g., 3) of CDR-L1,CDR-L2 and CDR-L3 of the 4A10 V_(L) (e.g., SEQ ID NO: 7); e.g., whereinthe CDRs comprise the amino acid sequences set forth in SEQ ID NOs: 8(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions), 9 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 10(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody heavy chainvariable (V_(H)) domain comprising one or more (e.g., 3) of CDR-H1,CDR-H2 or CDR-H3 of 19E8 V_(H) (e.g., SEQ ID NO: 12); e.g., wherein theCDRs comprise the amino acid sequences set forth in SEQ ID NOs: 13 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), 14 (or a variant thereof having 1, 2, 3, 4, 5,6, 7, 8, 9 or 10 point mutations and/or point deletions) and 15 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody light chainvariable (V_(L)) domain comprising one or more (e.g., 3) of CDR-L1,CDR-L2 and CDR-L3 of 19E8 V_(L) (e.g., SEQ ID NO: 17); e.g., wherein theCDRs comprise the amino acid sequences set forth in SEQ ID NOs: 18 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), 19 (or a variant thereof having 1, 2, 3, 4, 5,6, 7, 8, 9 or 10 point mutations and/or point deletions) and 20 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody heavy chainvariable (V_(H)) domain comprising one or more (e.g., 3) of CDR-H1,CDR-H2 or CDR-H3 of 11C9 V_(H) (e.g., SEQ ID NO: 22); e.g., wherein theCDRs comprise the amino acid sequences set forth in SEQ ID NOs: 23 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), 24 (or a variant thereof having 1, 2, 3, 4, 5,6, 7, 8, 9 or 10 point mutations and/or point deletions) and 25 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody light chainvariable (V_(L)) domain comprising one or more (e.g., 3) of CDR-L1,CDR-L2 and CDR-L3 of the 11C9 V_(L) (e.g., SEQ ID NO: 27); e.g., whereinthe CDRs comprise the amino acid sequences set forth in SEQ ID NOs: 28(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions), 29 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 30(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody heavy chainvariable (V_(H)) domain comprising one or more (e.g., 3) of CDR-H1,CDR-H2 or CDR-H3 of 22D2 V_(H) (e.g., SEQ ID NO: 32, 106, 108, 110, 112,114, 116, 118, 120 or 122); e.g., wherein the CDRs comprise the aminoacid sequences set forth in SEQ ID NOs: 33 (or a variant thereof having1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions),34, 446, 454, 455, 456, 457, or 458 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 35(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions), respectively.

The anti-LAG3 antibodies or antigen-binding fragments thereof of thepresent invention can comprise at least one antibody light chainvariable (V_(L)) domain comprising one or more (e.g., 3) of CDR-L1,CDR-L2 and CDR-L3 of 22D2 V_(L) (e.g., SEQ ID NO: 37 or 126); e.g.,wherein the CDRs comprise the amino acid sequences set forth in SEQ IDNOs: 38 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10point mutations and/or point deletions), 39 (or a variant thereof having1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)and 40 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10point mutations and/or point deletions), respectively.

The present invention provides an anti-LAG3 antibody or antigen-bindingfragment thereof that comprises:

-   -   the 4A10 CDR-H1, CDR-H2 and CDR-H3; and the 4A10 CDR-L1, CDR-L2        and CDR-L3;    -   the 11C9 CDR-H1, CDR-H2 and CDR-H3; and the 11C9 CDR-L1, CDR-L2        and CDR-L3;    -   the 19E8 CDR-H1, CDR-H2 and CDR-H3; and the 19E8 CDR-L1, CDR-L2        and CDR-L3; or    -   the 22D2 CDR-H1, CDR-H2 and CDR-H3; and the 22D2 CDR-L1, CDR-L2        and CDR-L3; wherein the 4A10, 11C9, 19E8 and 22D2 CDRs may be        derived from the mouse or humanized 4A10, 11C9, 19E8 and 22D2        immunoglobulin chains, respectively, set forth herein, and        wherein, optionally, 1, 2, 3, 4, 5 or 6 of the CDRs are variants        of those set forth herein.

The present invention provides an anti-LAG3 antibody or antigen-bindingfragment thereof that comprises an antibody light chain variable (V_(L))domain comprising a CDR-L1, CDR-L2 and CDR-L3 of the 4A10 V_(L) (e.g.,SEQ ID NOs: 8 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 point mutations and/or point deletions), 9 (or a variant thereofhaving 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or pointdeletions) and 10 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 point mutations and/or point deletions)); and an antibody heavychain variable (V_(H)) domain comprising a CDR-H1, CDR-H2 and CDR-H3 ofthe 4A10 V_(H) (e.g., SEQ ID NOs: 3 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 4 (ora variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions) and 5 (or a variant thereof having 1, 2, 3, 4,5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, an anti-LAG3 antibody or antigen-bindingfragment thereof comprises an antibody light chain variable (V_(L))domain comprising a CDR-L1, CDR-L2 and CDR-L3 of the 19E8 V_(L) (e.g.,SEQ ID NOs: 18 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 point mutations and/or point deletions), 19 (or a variant thereofhaving 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or pointdeletions) and 20 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 point mutations and/or point deletions)); and an antibody heavychain variable (V_(H)) domain comprising a CDR-H1, CDR-H2 and CDR-H3 ofthe 19E8 V_(H) (e.g., SEQ ID NOs: 13 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 14(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions) and 15 (or a variant thereof having 1,2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, an anti-LAG3 antibody or antigen-bindingfragment thereof comprises an antibody light chain variable (V_(L))domain comprising a CDR-L1, CDR-L2 and CDR-L3 of the 11C9 V_(L) (e.g.,SEQ ID NOs: 28 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 point mutations and/or point deletions), 29 (or a variant thereofhaving 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or pointdeletions) and 30 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 point mutations and/or point deletions)); and an antibody heavychain variable (V_(H)) domain comprising a CDR-H1, CDR-H2 and CDR-H3 ofthe 11C9 V_(H) (e.g., SEQ ID NOs: 23 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 24(or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 pointmutations and/or point deletions) and 25 (or a variant thereof having 1,2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions)).

In a further embodiment, an anti-LAG3 antibody or antigen-bindingfragment thereof comprises an antibody light chain variable (V_(L))domain comprising a CDR-L1, CDR-L2 and CDR-L3 of the 22D2 V_(L) (e.g.,SEQ ID NOs: 38 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or10 point mutations and/or point deletions), 39 (or a variant thereofhaving 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or pointdeletions) and 40 (or a variant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 point mutations and/or point deletions)); and an antibody heavychain variable (V_(H)) domain comprising a CDR-H1, CDR-H2 and CDR-H3 ofthe 22D2 V_(H) (e.g., SEQ ID NOs: 33 (or a variant thereof having 1, 2,3, 4, 5, 6, 7, 8, 9 or 10 point mutations and/or point deletions), 34,446, 454, 455, 456, 457 or 458 (ora variant thereof having 1, 2, 3, 4,5, 6, 7, 8, 9 or 10 point mutations and/or point deletions) and 35 (or avariant thereof having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 point mutationsand/or point deletions)).

In a further embodiment, the antibody is a humanized antagonistanti-LAG3 antibody. Examples of such humanized anti-LAG3 antibodiesinclude, but are not limited to, those comprising CDR-L1, CDR-L2 andCDR-L3 of 4A10; and CDR-H1, CDR-H2 and CDR-H3 of 4A10.

In a further embodiment, the antibody is a humanized antagonistanti-LAG3 antibody. Examples of such humanized anti-LAG3 antibodiesinclude, but are not limited to, those comprising CDR-L1, CDR-L2 andCDR-L3 of 19E8; and CDR-H1, CDR-H2 and CDR-H3 of 19E8.

In a further embodiment, the antibody is a humanized antagonistanti-LAG3 antibody. Examples of such humanized anti-LAG3 antibodiesinclude, but are not limited to, those comprising CDR-L1, CDR-L2 andCDR-L3 of 11C9; and CDR-H1, CDR-H2 and CDR-H3 of 11C9.

In a further embodiment, the antibody is a humanized antagonistanti-LAG3 antibody. Examples of such humanized anti-LAG3 antibodiesinclude, but are not limited to, those comprising CDR-L1, CDR-L2 andCDR-L3 of 22D2; and CDR-H1, CDR-H2 and CDR-H3 of 22D2; for example, Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 or Ab9.

The present invention provides an anti-LAG3 antibody or antigen-bindingfragment thereof or an immunoglobulin polypeptide that comprises:

-   -   the mature 4A10 V_(L) immunoglobulin domain and/or the mature        4A10 V_(H) domain;    -   the mature 19E8 V_(L) immunoglobulin domain and/or the mature        19E8 V_(H) domain;    -   the mature 11C9 V_(L) immunoglobulin domain and/or the mature        11C9 V_(H) domain; and/or    -   the mature 22D2 V_(L) immunoglobulin domain and/or the mature        22D2 V_(H) domain; wherein the 4A10, 19E8, 11C9 and 22D2 V_(L)        or V_(H) domain is a mouse or humanized 4A10, 19E8, 11C9 and        22D2 V_(L) or V_(H) domain set forth herein, and wherein,        optionally, the V_(L) and/or V_(H) is a variant of a V_(L) or        V_(H) set forth herein (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,        Ab8 or Ab9).

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(L) domainof 4A10, 19E8, 11C9 or 22D2 wherein the V_(L) domain comprises the aminoacid sequence of SEQ ID NO: 7, 17, 27, 37, 57, 59, 61, 63, 65, 101, 126,130, 132, 136, 138, 208, 210, 224, 226, 228, 230, 232, 241, 257, 259,261, 263, 351, 369, 371, 373, 375, 401, 403, 405, 426, 427, 450-453 or459-461 or a variant thereof

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(H) domainof 4A10, 19E8, 11C9 or 22D2 wherein the V_(H) domain comprises the aminoacid sequence of SEQ ID NO: 2, 12, 22, 32, 45, 47, 49, 51, 53, 55, 67,69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 103,106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 128, 134, 140, 142,144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170,172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198,200, 202, 204, 206, 212, 214, 216, 218, 220, 222, 234, 235, 237, 239,243, 245, 247, 249, 251, 253, 255, 265, 267, 269, 271, 273, 275, 277,279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333,335, 337, 339, 341, 343, 345, 347, 349, 353, 355, 357, 359, 361, 363,365, 367, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399,406-419, 434-442, 448, 449, 462, 463 or 464; or a variant thereof.

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(L) domainof 4A10 (e.g., amino acids 21-132 of SEQ ID NO: 7 or a variant thereof)and the mature V_(H) domain of 4A10 (e.g., amino acids 20-136 of SEQ IDNO: 2 or a variant thereof).

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(L) domainof 19E8 (e.g., amino acids 21-128 of SEQ ID NO: 17 or a variant thereof)and the mature V_(H) domain of 19E8 (e.g., amino acids 20-138 of SEQ IDNO: 12 or a variant thereof).

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(L) domainof 11C9 (e.g., amino acids 21-128 of SEQ ID NO: 27 or a variant thereof)and the mature V_(H) domain of 11C9 (e.g., amino acids 20-142 of SEQ IDNO: 22 or a variant thereof).

The present invention further provides an anti-LAG3 antibody orantigen-binding fragment thereof that comprises the mature V_(L) domainof 22D2 (e.g., amino acids 21-131 of SEQ ID NO: 37 or 126 or a variantthereof) and the mature V_(H) domain of 22D2 (e.g., amino acids 21-138or 21-131 of SEQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118, 120 or122 or a variant thereof).

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 2 or amino acids 20-136 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 7 or amino acids 21-132 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 12 or amino acids 20-138 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 17 or amino acids 21-128 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 22 or amino acids 20-142 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 27 or amino acids 21-128 thereof or avariant thereof; or any polynucleotide encoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 32, 106, 108, 110, 112, 114, 116, 118,120 or 122 or a mature fragment thereof, e.g., comprising amino acids20-138 or 20-131 thereof; or a variant thereof; or any polynucleotideencoding such a polypeptide.

The invention also provides polypeptides comprising the amino acidsequence set forth in SEQ ID NO: 37 or 126 or a mature fragment thereof,e.g., comprising amino acids 21-131 thereof or a variant thereof; or anypolynucleotide encoding such a polypeptide.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-H1, CDR-H2, and CDR-H3 of aV_(H) domain comprising SEQ ID NO: 2 (e.g., SEQ ID NOs: 3-5); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-L1, CDR-L2, and CDR-L3 of aV_(L) domain comprising SEQ ID NO: 7 (e.g., SEQ ID NOs: 8-10); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-H1, CDR-H2, and CDR-H3 of aV_(H) domain comprising SEQ ID NO: 12 (e.g., SEQ ID NOs: 13-15); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-L1, CDR-L2, and CDR-L3 of aV_(L) domain comprising SEQ ID NO: 17 (e.g., SEQ ID NOs: 18-20); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-H1, CDR-H2, and CDR-H3 of aV_(H) domain comprising SEQ ID NO: 22 (e.g., SEQ ID NOs: 23-25); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-L1, CDR-L2, and CDR-L3 of aV_(L) domain comprising SEQ ID NO: 27 (e.g., SEQ ID NOs: 28-30); or anypolynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3 ofsuch CDRs are variants of the sequence set forth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-H1, CDR-H2, and CDR-H3 of aV_(H) domain comprising SEQ ID NO: 32, 106, 108, 110, 112, 114, 116,118, 120 or 122 (e.g., SEQ ID NOs: 33, 34 (or 446, 454, 455, 456, 457 or458) or 35); or any polynucleotide encoding such a polypeptide.Optionally, 1, 2 or 3 of such CDRs are variants of the sequence setforth herein.

The invention also provides polypeptides (e.g., a humanizedimmunoglobulin chain) comprising the CDR-L1, CDR-L2, and CDR-L3 of aV_(L) domain comprising SEQ ID NO: 37 or 126 (e.g., SEQ ID NOs: 38-40);or any polynucleotide encoding such a polypeptide. Optionally, 1, 2 or 3of such CDRs are variants of the sequence set forth herein.

The present invention includes crystalline compositions of the anti-LAG3antibodies and antigen-binding fragments thereof of the presentinvention.

Polynucleotides

The present invention further comprises the polynucleotides encoding anyof the polypeptides or immunoglobulin chains of anti-LAG3 antibodies andantigen-binding fragments thereof disclosed herein (including variantsof the amino acid chains specifically set forth herein). For example,the present invention includes the polynucleotides described in SEQ IDNOs: 1, 6, 11, 16, 21, 26, 31, 36, 46, 48, 50, 52, 54, 56, 56, 58, 60,62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96,98, 100, 102, 104, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123,125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151,153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179,181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207,209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 236,238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264,266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292,294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320,322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348,350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376,378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402 or 404and variants thereof (e.g., comprising nucleotide sequences having atleast 70%, 80%, 90%, 95% or 99% BLAST sequence identity to suchnucleotide sequences (as discussed above)); and polynucleotides encodingthe amino acids described therein, e.g., in SEQ ID NOs: 2, 3, 4, 5, 7,8, 9, 10, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 27, 28, 29,30, 32, 33, 34, 35, 37, 38, 39, 40, 45, 47, 49, 51, 53, 55, 57, 59, 61,63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97,99, 101, 103, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126,128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154,156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182,184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210,212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 235, 237,239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265,267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293,295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321,323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349,351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377,379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405,406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419,426, 427, 434, 435, 436, 437, 438, 439, 440, 441, 442, 446, 448, 449,451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463 or 464.The scope of the present invention also includes variant polynucleotidesthat hybridize to any of such polynucleotides.

Moreover, the present invention includes anti-LAG3 antibodies andantigen-binding fragments thereof comprising immunoglobulin heavy andlight chains (e.g., variable regions thereof) and/or heavy and lightchain CDRs encoded by the polynucleotides set forth herein.

For example, the present invention includes anti-LAG3 antibodies andantigen-binding fragments thereof comprising a heavy chainimmunoglobulin encoded by a polynucleotide comprising the nucleotidesequence set forth in SEQ ID NO: 107 (or encoding a variable domainthereof) and a light chain immunoglobulin encoded by the nucleotidesequence set forth in SEQ ID NO: 125 (or encoding a variable domainthereof). For example, the present invention also includes anti-LAG3antibodies and antigen-binding fragments thereof comprising a heavychain immunoglobulin encoded by a polynucleotide comprising thenucleotide sequence set forth in SEQ ID NO: 115 (or encoding a variabledomain thereof) and a light chain immunoglobulin encoded by thenucleotide sequence set forth in SEQ ID NO: 125 (or encoding a variabledomain thereof).

The present invention provides polynucleotide encoding the

-   -   4A10 V_(L) or a mature fragment thereof;    -   4A10 V_(H) or a mature fragment thereof;    -   19E8 V_(L) or a mature fragment thereof;    -   19E8 V_(H) or a mature fragment thereof;    -   11C9 V_(L) or a mature fragment thereof    -   11C9 V_(H) or a mature fragment thereof;    -   22D2 V_(L) or a mature fragment thereof; and/or    -   22D2 V_(H) or a mature fragment thereof; wherein the 4A10, 19E8,        11C9 and 22D2 V_(L) or V_(H) domain is a mouse or humanized        4A10, 19E8, 11C9 and 22D2 V_(L) or V_(H) domain set forth        herein, and wherein, optionally, the V_(L) and/or V_(H) is a        variant of a V_(L) or V_(H) set forth herein.

The invention also provides polynucleotide comprising the nucleotidesequence set forth in SEQ ID NO: 1 or nucleotide 58-408 thereof; or avariant thereof.

The invention also provides polynucleotide comprising the nucleotidesequence set forth in SEQ ID NO: 6 or nucleotide 61-396 thereof; or avariant thereof.

The invention also provides polynucleotide comprising the nucleotidesequence set forth in SEQ ID NO:11 or nucleotide 58-414 thereof; or avariant thereof.

The invention also provides polynucleotides comprising the nucleotidesequence set forth in SEQ ID NO: 16 or nucleotide 61-384 thereof; or avariant thereof.

The invention also provides polynucleotides comprising the nucleotidesequence set forth in SEQ ID NO: 21 or nucleotide 58-426 thereof; or avariant thereof.

The invention also provides polynucleotides comprising the nucleotidesequence set forth in SEQ ID NO: 26 or nucleotide 61-384 thereof; or avariant thereof.

The invention also provides polynucleotides comprising the nucleotidesequence set forth in SEQ ID NO: 31 or nucleotide 61-447 thereof; or avariant thereof.

The invention also provides polynucleotides comprising the nucleotidesequence set forth in SEQ ID NO: 36 or nucleotide 61-547 thereof; or avariant thereof.

Variant polynucleotides set forth herein include those that hybridizeunder low, moderate or high stringency conditions to the polynucleotidesset forth herein or to polynucleotides that encode the polypeptides setforth herein, and encode immunoglobulin chains of anti-LAG3 antibodiesor antigen-binding fragments thereof which maintain the ability tospecifically bind to LAG3 (human and/or cynomolgous monkey, e.g., Macacafascicularis or Macaca mulatta). A first polynucleotide molecule is“hybridizable” to a second polynucleotide molecule when a singlestranded form of the first polynucleotide molecule can anneal to thesecond polynucleotide molecule under the appropriate conditions oftemperature and solution ionic strength (see Sambrook, et al., supra).The conditions of temperature and ionic strength determine the“stringency” of the hybridization. Typical low stringency hybridizationconditions include 55° C., 5×SSC, 0.1% SDS and no formamide; or 30%formamide, 5×SSC, 0.5% SDS at 42° C. Typical moderate stringencyhybridization conditions are 40% formamide, with 5× or 6×SSC and 0.1%SDS at 42° C. High stringency hybridization conditions are 50%formamide, 5× or 6×SSC at 42° C. or, optionally, at a higher temperature(e.g., 57° C., 59° C., 60° C., 62° C., 63° C., 65° C. or 68° C.). Ingeneral, SSC is 0.15M NaCl and 0.015M Na-citrate. Hybridization requiresthat the two polynucleotide contain complementary sequences, although,depending on the stringency of the hybridization, mismatches betweenbases are possible. The appropriate stringency for hybridizingpolynucleotides depends on the length of the polynucleotides and thedegree of complementation, variables well known in the art. The greaterthe degree of similarity or homology between two nucleotide sequences,the higher the stringency under which the nucleic acids may hybridize.For hybrids of greater than 100 nucleotides in length, equations forcalculating the melting temperature have been derived (see Sambrook, etal., supra, 9.50-9.51). For hybridization with shorter polynucleotides,e.g., oligonucleotides, the position of mismatches becomes moreimportant, and the length of the oligonucleotide determines itsspecificity (see Sambrook, et al., supra, 11.7-11.8).

In another embodiment of the invention, an polynucleotide, for exampleDNA, encoding the immunoglobulin polypeptide chains of the anti-LAG3antibodies or antigen-binding fragments set forth herein forms part ofthe present invention. In one embodiment, the polynucleotide encodes atleast one mature immunoglobulin polypeptide light chain variable (V_(L))domain and at least one mature immunoglobulin polypeptide heavy chainvariable (V_(H)) domain, wherein the V_(L) domain comprises a CDR-L1,CDR-L2 and CDR-L3 having a sequence selected from SEQ ID NOs: 8-10,18-20 28-30 and 38-40, and the V_(H) domain comprises CDR-H1, CDR-H2 andCDR-H3 having a sequence selected from SEQ ID NOs: 3-5, 13-15, 23-25 and33-35. In one embodiment, the nucleic acid encodes the 4A10, 11C9, 19E8or 22D2 mature light chain variable region and/or the 4A10, 11C9, 19E8or 22D2 mature heavy chain variable region sequences. In someembodiments of the invention, the polynucleotide encodes both a lightchain and a heavy chain on a single polynucleotide molecule, and, inother embodiments of the invention, the light and heavy chains areencoded on separate polynucleotide molecules, e.g., in separate orcommon host cells. In another embodiment the polynucleotides furtherencodes a signal sequence.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin light chain variable (V_(L)) domain comprising theCDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 7. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin light chain variable (V_(L)) domain comprising theCDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 17. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin light chain variable (V_(L)) domain comprising theCDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 27. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin light chain variable (V_(L)) domain comprising theCDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 37 or 126. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin heavy chain variable (V_(H)) domain comprising theCDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 2. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin heavy chain variable (V_(H)) domain comprising theCDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 12. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin heavy chain variable (V_(H)) domain comprising theCDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 22. Variants of suchpolynucleotides are also part of the present invention.

In one embodiment of the invention, the polynucleotide encodes a matureimmunoglobulin heavy chain variable (V_(H)) domain comprising theCDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 32, 106, 108, 110, 112, 114,116, 118, 120 or 122. Variants of such polynucleotides are also part ofthe present invention.

In one embodiment of the invention, the polynucleotide encodes theimmunoglobulin light chain variable (V_(L)) domain of SEQ ID NO: 7, 17,27 and/or 37. Variants of such polynucleotides are also part of thepresent invention.

In one embodiment of the invention, the polynucleotide encodes theimmunoglobulin heavy chain variable (V_(H)) domain of SEQ ID NO: 2, 12,22, 32, 106, 108, 110, 112, 114, 116, 118, 120 and/or 122. Variants ofsuch polynucleotides are also part of the present invention.

This present invention also provides vectors, e.g., expression vectors,such as plasmids, comprising the polynucleotides of the invention(sequences set forth herein and variants thereof, e.g., SEQ ID NO: 125,105, 107, 109, 111, 113, 115, 117, 119 and/or 121), wherein thepolynucleotide is operably linked to control sequences that arerecognized by a host cell when the host cell is transfected with thevector. Also provided are host cells comprising a polynucleotide (e.g.,integrated into the genome, e.g., a chromosome, of the host cell) orvector of the present invention and methods for producing the antibodyor antigen-binding fragment thereof or polypeptide disclosed hereincomprising culturing a host cell harboring an expression vector orpolynucleotide encoding the immunoglobulin chains of the antibody orantigen-binding fragment thereof in culture medium, and isolating theantigen or antigen-binding fragment thereof from the host cell orculture medium.

Binding Affinity

By way of example, and not limitation, the anti-LAG3 antibodies andantigen-binding fragments thereof disclosed herein bind human and/orcynomolgous monkey, e.g., Macaca fascicularis or Macaca mulatta LAG3,e.g., with a K_(D) value of at least about 100 nM (1×10⁻⁷M); at leastabout 10 nM; or at least about 1 nM. In further embodiments, theantibodies have K_(D) values of at least about 200 pM (2×10⁻¹⁰ M), 100pM, 50 pM, 20 pM, 10 pM, 5 pM or even 2 pM. For example, the K_(D) isabout 2.77×10⁻¹² M, 1.47×10⁻¹¹ M, 1.47×10⁻⁰⁹ M, or 9.03×10⁻¹¹ M; or ahigher affinity. In an embodiment of the invention, the K_(D) is asmeasured in a KinExA assay or similar kinetic exclusion assay. See e.g.,Darling et al. Assay and Drug Dev. Tech. 2(6): 647-657 (2004).

Methods of Making Antibodies and Antigen-binding Fragments Thereof

Hybridoma cells that produce parental (e.g., mouse) monoclonal anti-LAG3antibodies or antigen-binding fragments thereof discussed herein may beproduced by methods which are commonly known in the art. Such isolatedhybridomas are part of the present invention. These methods include, butare not limited to, the hybridoma technique originally developed byKohler, et al., (1975) (Nature 256:495-497), as well as the triomatechnique (Hering, et al., (1988) Biomed. Biochim. Acta. 47:211-216 andHagiwara, et al., (1993) Hum. Antibod. Hybridomas 4:15), the humanB-cell hybridoma technique (Kozbor, et al., (1983) Immunology Today 4:72and Cote, et al., (1983) Proc. Natl. Acad. Sci. U.S.A 80:2026-2030), theEBV-hybridoma technique (Cole, et al., in Monoclonal Antibodies andCancer Therapy, Alan R. Liss, Inc., pp. 77-96, 1985), and electric fieldbased electrofusion using a Cyto Pulse large chamber cull fusionelectroporator (Cyto Pulse Sciences, Inc., Glen Burnie, Md.).Preferably, mouse splenocytes are isolated and fused with PEG or byelectrofusion to a mouse myeloma cell line based upon standardprotocols. The resulting hybridomas may then be screened for theproduction of antigen-specific antibodies. For example, single cellsuspensions of splenic lymphocytes from immunized mice may by fused toone-sixth the number of P3X63-Ag8.653 nonsecreting mouse myeloma cells(ATCC, CRL 1580) with 50% PEG. Cells may be plated at approximately2×10⁵ cells/mL in a flat bottom microtiter plate, followed by a two weekincubation in selective medium containing 20% fetal Clone Serum, 18%“653” conditioned media, 5% origen (IGEN), 4 mM L-glutamine, 1 mML-glutamine, 1 mM sodium pyruvate, 5 mM HEPES, 0.055 mM2-mercaptoethanol, 50 units/ml penicillin, 50 mg/ml streptomycin, 50mg/ml gentamycin and 1× HAT (Sigma; the HAT is added 24 hours after thefusion). After two weeks, cells may be cultured in medium in which theHAT is replaced with HT. Individual wells may then be screened by ELISAfor anti-LAG3 monoclonal IgG antibodies. Once extensive hybridoma growthoccurs, medium can be observed usually after 10-14 days. The antibodysecreting hybridomas may be replated, screened again, and if stillpositive for human IgG, anti-LAG3 monoclonal antibodies, can besubcloned at least twice by limiting dilution. The stable subclones maythen be cultured in vitro to generate small amounts of antibody intissue culture medium for characterization.

Thus, the present invention includes methods for making an anti-LAG3antibody or antigen-binding fragment thereof of the present inventioncomprising culturing a hybridoma cell that expresses the antibody orfragment under condition favorable to such expression and, optionally,isolating the antibody or fragment from the hybridoma.

The anti-LAG3 antibodies disclosed herein may also be producedrecombinantly (e.g., in an E. coli/T7 expression system). In thisembodiment, nucleic acids encoding the anti-LAG3 antibody immunoglobulinmolecules of the invention (e.g., V_(H) or V_(L); e.g., any one or moreof SEQ ID NO: 125, 105, 107, 109, 111, 113, 115, 117, 119 and/or 121)may be inserted into a pET-based plasmid and expressed in the E. coli/T7system. For example, the present invention includes methods forexpressing an antibody or antigen-binding fragment thereof orimmunoglobulin chain thereof in a host cell (e.g., bacterial host cellsuch as E. coli such as BL21 or BL21DE3) comprising expressing T7 RNApolymerase in the cell which also includes a polynucleotide encoding animmunoglobulin chain that is operably linked to a T7 promoter. Forexample, in an embodiment of the invention, a bacterial host cell, suchas a E. coli, includes a polynucleotide encoding the T7 RNA polymerasegene operably linked to a lac promoter and expression of the polymeraseand the chain is induced by incubation of the host cell with IPTG(isopropyl-beta-D-thiogalactopyranoside).

There are several methods by which to produce recombinant antibodieswhich are known in the art. One example of a method for recombinantproduction of antibodies is disclosed in U.S. Pat. No. 4,816,567.

Transformation can be by any known method for introducingpolynucleotides into a host cell. Methods for introduction ofheterologous polynucleotides into mammalian cells are well known in theart and include dextran-mediated transfection, calcium phosphateprecipitation, polybrene-mediated transfection, protoplast fusion,electroporation, encapsulation of the polynucleotide(s) in liposomes,biolistic injection and direct microinjection of the DNA into nuclei. Inaddition, nucleic acid molecules may be introduced into mammalian cellsby viral vectors. Methods of transforming cells are well known in theart. See, for example, U.S. Pat. Nos. 4,399,216; 4,912,040; 4,740,461and 4,959,455.

Thus, the present invention includes recombinant methods for making ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention, or an immunoglobulin chain thereof, comprising (i)introducing a polynucleotide (e.g., any one or more of SEQ ID NO: 125,105, 107, 109, 111, 113, 115, 117, 119 and/or 121) encoding one or moreimmunoglobulin chains of the antibody or fragment (e.g., heavy chainimmunoglobulin of 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9 and/or light chain immunoglobulin of4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9), for example, wherein the polynucleotide is in a vectorand/or is operably linked to a promoter; (ii) culturing the host cell(e.g., CHO or Pichia or Pichia pastoris) under condition favorable toexpression of the polynucleotide and, (iii) optionally, isolating theantibody or fragment or chain from the host cell and/or medium in whichthe host cell is grown. When making an antibody or antigen-bindingfragment comprising more than one immunoglobulin chain, e.g., anantibody that comprises two heavy immunoglobulin chains and two lightimmunoglobulin chains, co-expression of the chains in a single host cellleads to association of the chains, e.g., in the cell or on the cellsurface or outside the cell if such chains are secreted, so as to formthe antibody or antigen-binding fragment molecule. The methods includethose wherein only a heavy immunoglobulin chain or only a lightimmunoglobulin chain (e.g., any of those discussed herein includingmature fragments and/or variable domains thereof) is expressed. Suchchains are useful, for example, as intermediates in the expression of anantibody or antigen-binding fragment that includes such a chain. Forexample, the present invention also includes anti-LAG3 antibodies andantigen-binding fragments thereof comprising a heavy chainimmunoglobulin (or variable domain thereof or comprising the CDRsthereof) encoded by a polynucleotide comprising the nucleotide sequenceset forth in SEQ ID NO: 115 (or encoding a variable domain thereof) anda light chain immunoglobulin (or variable domain thereof or comprisingthe CDRs thereof) encoded by the nucleotide sequence set forth in SEQ IDNO: 125 (or encoding a variable domain thereof) which are the product ofsuch production methods, and, optionally, the purification methods setforth herein.

Anti-LAG3 antibodies can also be synthesized by any of the methods setforth in U.S. Pat. No. 6,331,415.

Eukaryotic and prokaryotic host cells, including mammalian cells ashosts for expression of the anti-LAG3 antibodies or fragments orimmunoglobulin chains disclosed herein are well known in the art andinclude many immortalized cell lines available from the American TypeCulture Collection (ATCC). These include, inter alia, Chinese hamsterovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK)cells, monkey kidney cells (COS), human hepatocellular carcinoma cells(e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells and a number ofother cell lines. Mammalian host cells include human, mouse, rat, dog,monkey, pig, goat, bovine, horse and hamster cells. Cell lines ofparticular preference are selected through determining which cell lineshave high expression levels. Other cell lines that may be used areinsect cell lines (e.g., Spodoptera frupperda or Trichoplusia ni),amphibian cells, bacterial cells, plant cells and fungal cells. Fungalcells include yeast and filamentous fungus cells including, for example,Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichiakoclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichialindneri), Pichia opuntiae, Pichia thermotolerans, Pichia salictaria,Pichia guercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica,Pichia sp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenulapolymorpha, Kluyveromyces sp., Kluyveromyces lactis, Candida albicans,Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichodermareesei, Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum,Fusarium venenatum, Physcomitrella patens and Neurospora crassa. Pichiasp., any Saccharomyces sp., Hansenula polymorpha, any Kluyveromyces sp.,Candida albicans, any Aspergillus sp., Trichoderma reesei, Chrysosporiumlucknowense, any Fusarium sp., Yarrowia lipolytica, and Neurosporacrassa.

Further, expression of antibodies and antigen-binding fragments thereofand immunoglobulin chains of the invention (or other moieties therefrom)from production cell lines can be enhanced using a number of knowntechniques. For example, the glutamine synthetase gene expression system(the GS system) is a common approach for enhancing expression undercertain conditions. The GS system is discussed in whole or part inconnection with European Pat. Nos. 0 216 846, 0 256 055, and 0 323 997and European Patent Application No. 89303964.4. Thus, in an embodimentof the invention, the mammalian host cells (e.g., CHO) lack a glutaminesynthetase gene and are grown in the absence of glutamine in the mediumwherein, however, the polynucleotide encoding the immunoglobulin chaincomprises a glutamine synthetase gene which complements the lack of thegene in the host cell.

The present invention includes methods for purifying an anti-LAG3antibody or antigen-binding fragment thereof of the present inventioncomprising introducing a sample (e.g., culture medium, cell lysate orcell lysate fraction, e.g., a soluble fraction of the lysate) comprisingthe antibody or fragment to a purification medium (e.g., cation-exchangemedium, anion-exchange medium, hydrophobic exchange medium, affinitypurification medium (e.g., protein-A, protein-G, protein-A/G,protein-L)) and either collecting purified antibody or fragment from theflow-through fraction of said sample that does not bind to the medium;or, discarding the flow-through fraction and eluting bound antibody orfragment from the medium and collecting the eluate. In an embodiment ofthe invention, the medium is in a column to which the sample is applied.In an embodiment of the invention, the purification method is conductedfollowing recombinant expression of the antibody or fragment in a hostcell, e.g., wherein the host cell is first lysed and, optionally, thelysate is purified of insoluble materials prior to purification on amedium; or wherein the antibody or fragment is secreted into the culturemedium by the host cell and the medium or a fraction thereof is appliedto the purification medium.

In general, glycoproteins produced in a particular cell line ortransgenic animal will have a glycosylation pattern that ischaracteristic for glycoproteins produced in the cell line or transgenicanimal. Therefore, the particular glycosylation pattern of an antibodywill depend on the particular cell line or transgenic animal used toproduce the antibody. However, all antibodies encoded by the nucleicacid molecules provided herein, or comprising the amino acid sequencesprovided herein, comprise the instant invention, independent of theglycosylation pattern the antibodies may have. Similarly, in particularembodiments, antibodies with a glycosylation pattern comprising onlynon-fucosylated N-glycans may be advantageous, because these antibodieshave been shown to typically exhibit more potent efficacy than theirfucosylated counterparts both in vitro and in vivo (See for example,Shinkawa et al., J. Biol. Chem. 278: 3466-3473 (2003); U.S. Pat. Nos.6,946,292 and 7,214,775). These antibodies with non-fucosylatedN-glycans are not likely to be immunogenic because their carbohydratestructures are a normal component of the population that exists in humanserum IgG.

The present invention includes anti-LAG3 antibodies and antigen-bindingfragments thereof (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) comprising N-linked glycansthat are typically added to immunoglobulins produced in Chinese hamsterovary cells (CHO N-linked glycans) or to engineered yeast cells(engineered yeast N-linked glycans), such as, for example, Pichiapastoris. For example, in an embodiment of the invention, the antibodyor antigen-binding fragment comprises one or more of the “engineeredyeast N-linked glycans” or “CHO N-linked glycans” that are set forth inFIG. 6 (e.g., G0 and/or G0-F and/or G1 and/or G1-F and/or and/or G2-Fand/or Man5). In an embodiment of the invention, the antibody orantigen-binding fragment comprises the engineered yeast N-linkedglycans, i.e., G0 and/or G1 and/or G2, optionally, further includingMan5. In an embodiment of the invention, the antibody or antigen-bindingfragment comprise the CHO N-linked glycans, i.e., G0-F, G1-F and G2-F,optionally, further including G0 and/or G1 and/or G2 and/or Man5. In anembodiment of the invention, about 80% to about 95% (e.g., about 80-90%,about 85%, about 90% or about 95%) of all N-linked glycans on theantibody or antigen-binding fragment immunoglobulin chains areengineered yeast N-linked glycans or CHO N-linked glycans. See Nett etal. Yeast. 28(3): 237-252 (2011); Hamilton et al. Science. 313(5792):1441-1443 (2006); Hamilton et al. Curr Opin Biotechnol. 18(5): 387-392(2007). For example, in an embodiment of the invention, an engineeredyeast cell is GFI5.0 or YGLY8316 or strains set forth in U.S. Pat. No.7,795,002 or Zha et al. Methods Mol Biol. 988:31-43 (2013). See alsointernational patent application publication no. WO2013/066765.

The present invention includes polyclonal anti-LAG3 antibodies andantigen-binding fragments thereof, e.g., a composition comprising aplurality of anti-LAG3 antibodies and fragments, which include one ormore of the anti-LAG3 antibodies or antigen-binding fragments thereof ofthe present invention (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 orAb9), and methods of use thereof. A polyclonal antibody is an antibodywhich was produced among or in the presence of one or more other,non-identical antibodies. In general, polyclonal antibodies are producedfrom collections of different B-lymphocytes, e.g., the B-lymphocyte ofan animal treated with an immunogen of interest, which produces apopulation of different antibodies but which are all directed to theimmunogen. Usually, polyclonal antibodies are obtained directly from animmunized animal, e.g., spleen, serum or ascites fluid.

The present invention includes “antagonist” anti-LAG3 antibodies andantigen-binding fragments thereof and methods of use thereof, e.g.,humanized, antagonist anti-LAG3 antibodies and fragments. An antagonistanti-LAG3 antibody or antigen-binding fragment thereof antagonizes anactivity of LAG3 (e.g., human LAG3) such as by inhibiting LAG3 bindingto MHC class II molecules; competing with MHC class II molecules forLAG3 binding; or when a cell or subject is contacted with the antibodyor fragment, a biological phenotype associated with LAG3 antagonism,such as stimulation of antigen-specific T-cell production of IL-2, isproduced.

The present invention includes bispecific and bifunctional antibodiesand antigen-binding fragments having a binding specificity for LAG3 andanother antigen such as, for example, PD-1 or PD-L1, and methods of usethereof. In an embodiment of the invention, the anti-PD1 chains comprisethe amino acid sequence of SEQ ID NOs: 41 and 42 or of SEQ ID NOs: 43and 44. A bispecific or bifunctional antibody is an artificial hybridantibody having two different heavy/light chain pairs and two differentbinding sites. Bispecific antibodies can be produced by a variety ofmethods including fusion of hybridomas or linking of Fab′ fragments.See, e.g., Songsivilai, et al., (1990) Clin. Exp. Immunol. 79: 315-321,Kostelny, et al., (1992) J Immunol. 148:1547-1553. In addition,bispecific antibodies may be formed as “diabodies” (Holliger, et al.,(1993) PNAS USA 90:6444-6448) or as “Janusins” (Traunecker, et al.,(1991) EMBO J. 10:3655-3659 and Traunecker, et al., (1992) Int. J.Cancer Suppl. 7:51-52).

The present invention further includes anti-LAG3 antigen-bindingfragments of the anti-LAG3 antibodies disclosed herein. The antibodyfragments include F(ab)₂ fragments, which may be produced by enzymaticcleavage of an IgG by, for example, pepsin. Fab fragments may beproduced by, for example, reduction of F(ab)₂ with dithiothreitol ormercaptoethylamine. A Fab fragment is a V_(L)-C_(L) chain appended to aV_(H)-C_(H1) chain by a disulfide bridge. A F(ab)₂ fragment is two Fabfragments which, in turn, are appended by two disulfide bridges. The Fabportion of an F(ab)₂ molecule includes a portion of the F_(c) regionbetween which disulfide bridges are located. An F_(V) fragment is aV_(L) or V_(H) region.

Immunoglobulins may be assigned to different classes depending on theamino acid sequences of the constant domain of their heavy chains. Thereare at least five major classes of immunoglobulins: IgA, IgD, IgE, IgGand IgM, and several of these may be further divided into subclasses(isotypes), e.g. IgG-1, IgG-2, IgG-3 and IgG-4; IgA-1 and IgA-2. Theinvention comprises anti-LAG3 antibodies and antigen-binding fragmentsof any of these classes or subclasses of antibodies.

In one embodiment, the anti-LAG3 antibody or antigen-binding fragmentcomprises a heavy chain constant region, e.g. a human constant region,such as γ, γ2, γ3, or γ4 human heavy chain constant region or a variantthereof. In another embodiment, the anti-LAG3 antibody orantigen-binding fragment comprises a light chain constant region, e.g. ahuman light chain constant region, such as lambda or kappa human lightchain region or variant thereof. By way of example, and not limitation,the human heavy chain constant region can be γ4 and the human lightchain constant region can be kappa. In an alternative embodiment, the Fcregion of the antibody is γ4 with a Ser228Pro mutation (Schuurman, J et.al., Mol. Immunol. 38: 1-8, 2001).

In some embodiments, different constant domains may be appended tohumanized V_(L) and V_(H) regions derived from the CDRs provided herein.For example, if a particular intended use of an antibody (or fragment)of the present invention were to call for altered effector functions, aheavy chain constant domain other than human IgG1 may be used, or hybridIgG1/IgG4 may be utilized.

Although human IgG1 antibodies provide for long half-life and foreffector functions, such as complement activation and antibody-dependentcellular cytotoxicity, such activities may not be desirable for all usesof the antibody. In such instances a human IgG4 constant domain, forexample, may be used. The present invention includes anti-LAG3antibodies and antigen-binding fragments thereof which comprise an IgG4constant domain, e.g., antagonist, humanized anti-LAG3 antibodies andfragments, and methods of use thereof. In one embodiment, the IgG4constant domain can differ from the native human IgG4 constant domain(Swiss-Prot Accession No. P01861.1) at a position corresponding toposition 228 in the EU system and position 241 in the KABAT system,where the native Ser108 is replaced with Pro, in order to prevent apotential inter-chain disulfide bond between Cys106 and Cys109(corresponding to positions Cys 226 and Cys 229 in the EU system andpositions Cys 239 and Cys 242 in the KABAT system) that could interferewith proper intra-chain disulfide bond formation. See Angal et al.(1993) Mol. Imunol. 30:105. In other instances, a modified IgG1 constantdomain which has been modified to increase half-life or reduce effectorfunction can be used.

Antibody Engineering

Further included are embodiments in which the anti-LAG3 antibodies andantigen-binding fragments thereof are engineered antibodies to includemodifications to framework residues within the variable domains of aparental (e.g., mouse) monoclonal antibody, e.g. to improve theproperties of the antibody or fragment. Typically, such frameworkmodifications are made to decrease the immunogenicity of the antibody orfragment. This is usually accomplished by replacing non-CDR residues inthe variable domains (i.e. framework residues) in a parental (e.g.rodent) antibody or fragment with analogous residues from the immunerepertoire of the species in which the antibody is to be used, e.g.human residues in the case of human therapeutics. Such an antibody orfragment is referred to as a “humanized” antibody or fragment. In somecases it is desirable to increase the affinity, or alter the specificityof an engineered (e.g. humanized) antibody. One approach is to“backmutate” one or more framework residues to the correspondinggermline sequence. More specifically, an antibody or fragment that hasundergone somatic mutation can contain framework residues that differfrom the germline sequence from which the antibody is derived. Suchresidues can be identified by comparing the antibody or fragmentframework sequences to the germline sequences from which the antibody orfragment is derived. Another approach is to revert to the originalparental (e.g., rodent) residue at one or more positions of theengineered (e.g. humanized) antibody, e.g. to restore binding affinitythat may have been lost in the process of replacing the frameworkresidues. (See, e.g., U.S. Pat. Nos. 5,693,762, 5,585,089 and5,530,101.)

For example, Table 2, below, shows regions where a framework regionamino acid position (using Kabat numbering system) differs from thegermline and how this position can be backmutated to the germline by theindicated substitutions:

TABLE 2 Exemplary Backmutations Framework Amino Acid PositionBackmutation-- Region (Kabat Numbering) examples AbA V_(H) 25 H25S AbAV_(H) 68 S68T AbA V_(H)  82a T82aT

Another type of framework modification involves mutating one or moreresidues within the framework region, or even within one or more CDRregions, to remove T cell epitopes to thereby reduce the potentialimmunogenicity of the antibody. This approach is also referred to as“deimmunization” and is described in further detail in U.S. Pat. No.7,125,689.

In particular embodiments, it will be desirable to change certain aminoacids containing exposed side-chains to another amino acid residue inorder to provide for greater chemical stability of the final antibody,as follows. Such changes in the antigen-binding region can alter thebinding to the antigen. The deamidation of asparagine may occur on N-Gor D-G sequences and result in the creation of an isoaspartic acidresidue that introduces a kink into the polypeptide chain and decreasesits stability (isoaspartic acid effect). In certain embodiments, theantibodies of the present disclosure do not contain asparagine isomerismsites.

For example, an asparagine (Asn) residue may be changed to Gln or Ala toreduce the potential for formation of isoaspartate at any Asn-Glysequences, particularly within a CDR. A similar problem may occur at aAsp-Gly sequence. Reissner and Aswad (2003) Cell. Mol. Life Sci.60:1281. Isoaspartate formation may debilitate or completely abrogatebinding of an antibody to its target antigen. See, Presta (2005) J.Allergy Clin. Immunol. 116:731 at 734. In one embodiment, the asparagineis changed to glutamine (Gln). It may also be desirable to alter anamino acid adjacent to an asparagine (Asn) or glutamine (Gln) residue toreduce the likelihood of deamidation, which occurs at greater rates whensmall amino acids occur adjacent to asparagine or glutamine. See,Bischoff & Kolbe (1994) J. Chromatog. 662:261. In addition, anymethionine residues (typically solvent exposed Met) in CDRs may bechanged to Lys, Leu, Ala, or Phe or other amino acids in order to reducethe possibility that the methionine sulfur would oxidize, which couldreduce antigen-binding affinity and also contribute to molecularheterogeneity in the final antibody preparation. Id. In one embodimentof the invention, the methionine is changed to alanine (Ala).Additionally, in order to prevent or minimize potential scissile Asn-Propeptide bonds, it may be desirable to alter any Asn-Pro combinationsfound in a CDR to Gln-Pro, Ala-Pro, or Asn-Ala. Antibodies with suchsubstitutions are subsequently screened to ensure that the substitutionsdo not decrease the affinity or specificity of the antibody for LAG3, orother desired biological activity to unacceptable levels.

TABLE 3 Exemplary stabilizing CDR variants CDR Residue StabilizingVariant Sequence Asn-Gly Gln-Gly, Ala-Gly, or Asn-Ala (N-G) (Q-G),(A-G), or (N-A) Asp-Gly Glu-Gly, Ala-Gly or Asp-Ala (D-G) (E-G), (A-G),or (D-A) Met (typically solvent exposed) Lys, Leu, Ala, or Phe (M) (K),(L), (A), or (F) Asn Gln or Ala (N) (Q) or (A) Asn-Pro Gln-Pro, Ala-Pro,or Asn-Ala (N-P) (Q-P), (A-P), or (N-A)

The immunoglobulin chains set forth above 4A10, 19E8, 11C9 and 22D2contain residues that are double-underscored. The scope of the presentinvention includes antibodies, antigen-binding fragments, polypeptidesand polynucleotides as discussed herein wherein any one or more of suchresidues are mutated to any other residue including, for example, thoseof the stabilizing variant sequences set forth above in Table 3.

Mouse anti-LAG3 antibodies and antigen-binding fragments can behumanized by various methods known in the art (see e.g., humanizationmethods set forth in WO2005/047326 or U.S. Pat. No. 7,846,443.). Forexample, in an embodiment of the invention, mouse anti-LAG3 antibodiesand fragments are humanized by a method wherein computer aided molecularmodeling is used for identifying CDR loops in non-human immunoglobulinchains. This identification is made based upon the three-dimensionalstructure of the immunoglobulin chain and the position of the loops inthe chain.

Human frameworks (obtained from the IMGD Database), into which thenon-human loops will be introduced, are selected based on best matches(by amino acid sequence comparison) with the non-human sequence both inthe frameworks and in the CDRs. Regarding the FR4 in the V_(H) domain,VJ regions, for the human germlines, are compared with the correspondingnon-human VJ regions; and, regarding FR4 in V_(L) domain, J-kappa andJ-Lambda regions, of human germline sequences, are compared with thecorresponding non-human J-Kappa and J-Lambda regions.

Proper three-dimensional orientation of the CDRs, which is critical tomaintaining antigen binding, depends, in part, on proper interfacingbetween the V_(H) and V_(L). Thus, the molecular models are constructedand used for identifying residues in the V_(L)-V_(H) interface as wellas for identifying residues that can potentially alter the CDRconformations and hence binding to antigen. If necessary, mutations inthe immunoglobulin chain may be introduced so as to achieve desirableproperties e.g., antigen binding.

Developability Filters are established through the use of molecularmodeling techniques. Developability Filters are criteria used forfiltering features out of the final immunoglobulin chain so as to avoidunwanted effects. Molecular models are further used to identify solventexposed amino acids that can result in unwanted effects such asglycosylation, deamidation and oxidation. Such effects on the antibodycan lead to changes in the antibody conformation and hence function.Such problems can occur, for example, during scale-up or over aprolonged period of time when exposed to extreme chemical/physicalenvironments. Again, if necessary, mutations in the chains can beintroduced so as achieve the desired properties.

The Developability Filters are typically introduced early on in thedesign stage of the humanized chains to eliminate/minimize thesepotential problems. Humanized antibodies are further subjected to designcriteria, such as good expressibility and desirable isoelectric points.

Antibody Engineering of the Fc Region

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be engineered to includemodifications within the Fc region, typically to alter one or morefunctional properties of the antibody, such as serum half-life,complement fixation, Fc receptor binding, and/or effector function(e.g., antigen-dependent cellular cytotoxicity). Furthermore, theantibodies and antigen-binding fragments thereof (e.g., 4A10, 19E8, 11C9and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9)can be chemically modified (e.g., one or more chemical moieties can beattached to the antibody) or be modified to alter its glycosylation,again to alter one or more functional properties of the antibody orfragment. Each of these embodiments is described in further detailbelow. The numbering of residues in the Fc region is that of the EUindex of Kabat. Any such anti-LAG3 antibody or antigen-binding fragmentthereof having the modifications (e.g., Fc modifications) and/oralterations discussed herein are part of the present invention.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) also include antibodies and fragmentswith modified (or blocked) Fc regions to provide altered effectorfunctions. See, e.g., U.S. Pat. No. 5,624,821; WO2003/086310;WO2005/120571; WO2006/0057702. Such modifications can be used to enhanceor suppress various reactions of the immune system, with possiblebeneficial effects in diagnosis and therapy. Alterations of the Fcregion include amino acid changes (substitutions, deletions andinsertions), glycosylation or deglycosylation, and adding multiple Fc.Changes to the Fc can also alter the half-life of antibodies intherapeutic antibodies, enabling less frequent dosing and thus increasedconvenience and decreased use of material. See Presta (2005) J. AllergyClin. Immunol. 116:731 at 734-35.

In one embodiment, the anti-LAG3 antibody (e.g., humanized antibodiessuch as antagonist humanized antibodies) or antigen-binding fragment(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) is an IgG4 isotype antibody or fragment comprisinga serine to proline mutation at a position corresponding to position 228(S228P; EU index) in the hinge region of the heavy chain constantregion. This mutation has been reported to abolish the heterogeneity ofinter-heavy chain disulfide bridges in the hinge region (Angal et al.supra; position 241 is based on the Kabat numbering system).

In one embodiment of the invention, the hinge region of CH1 is modifiedsuch that the number of cysteine residues in the hinge region isincreased or decreased. This approach is described further in U.S. Pat.No. 5,677,425. The number of cysteine residues in the hinge region ofCH1 is altered, for example, to facilitate assembly of the light andheavy chains or to increase or decrease the stability of the antibody.

In another embodiment, the Fc hinge region of an anti-LAG3 antibody orantigen-binding fragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is mutated to decrease thebiological half-life of the antibody or fragment. More specifically, oneor more amino acid mutations are introduced into the CH2-CH3 domaininterface region of the Fc-hinge fragment such that the antibody orfragment has impaired Staphylococcyl protein A (SpA) binding relative tonative Fc-hinge domain SpA binding. This approach is described infurther detail in U.S. Pat. No. 6,165,745.

In another embodiment, the anti-LAG3 antibody (e.g., humanizedantibodies such as antagonist humanized antibodies) or antigen-bindingfragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) is modified to increase its biologicalhalf-life. Various approaches are possible. For example, one or more ofthe following mutations can be introduced: T252L, T254S, T256F, asdescribed in U.S. Pat. No. 6,277,375. Alternatively, to increase thebiological half-life, the antibody can be altered within the CH1 or CLregion to contain a salvage receptor binding epitope taken from twoloops of a CH2 domain of an Fc region of an IgG, as described in U.S.Pat. Nos. 5,869,046 and 6,121,022.

In yet other embodiments, the Fc region is altered by replacing at leastone amino acid residue with a different amino acid residue to alter theeffector function(s) of the anti-LAG3 antibody or antigen-bindingfragment. For example, one or more amino acids selected from amino acidresidues 234, 235, 236, 237, 297, 318, 320 and 322 can be replaced witha different amino acid residue such that the antibody has an alteredaffinity for an effector ligand but retains the antigen-binding abilityof the parent antibody. The effector ligand to which affinity is alteredcan be, for example, an Fc receptor or the C1 component of complement.This approach is described in further detail in U.S. Pat. Nos. 5,624,821and 5,648,260.

In another example, one or more amino acids selected from amino acidresidues 329, 331 and 322 can be replaced with a different amino acidresidue such that the anti-LAG3 antibody has altered C1q binding and/orreduced or abolished complement dependent cytotoxicity (CDC). Thisapproach is described in further detail in U.S. Pat. No. 6,194,551.

In another example, one or more amino acid residues within amino acidpositions 231 and 239 are altered to thereby alter the ability of theanti-LAG3 antibody or antigen-binding fragment thereof to fixcomplement. This approach is described further in PCT Publication WO94/29351.

In yet another example, the Fc region is modified to decrease theability of the anti-LAG3 antibody (e.g., humanized antibodies such asantagonist humanized antibodies) or antigen-binding fragment (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to mediate antibody dependent cellular cytotoxicity(ADCC) and/or to decrease the affinity of the antibody or fragment foran Fcγ receptor by modifying one or more amino acids at the followingpositions: 238, 239, 243, 248, 249, 252, 254, 255, 256, 258, 264, 265,267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290, 292,293, 294, 295, 296, 298, 301, 303, 305, 307, 309, 312, 315, 320, 322,324, 326, 327, 329, 330, 331, 333, 334, 335, 337, 338, 340, 360, 373,376, 378, 382, 388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or439. This approach is described further in PCT Publication WO 00/42072.Moreover, the binding sites on human IgG1 for FcγR1, FcγRII, FcγRIII andFcRn have been mapped and variants with improved binding have beendescribed (see Shields et al. (2001) J. Biol. Chem. 276:6591-6604).

In one embodiment of the invention, the Fc region is modified todecrease the ability of the anti-LAG3 antibody (e.g., humanizedantibodies such as antagonist humanized antibodies) or antigen-bindingfragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to mediate effector function and/or toincrease anti-inflammatory properties by modifying residues 243 and 264.In one embodiment, the Fc region of the antibody or fragment is modifiedby changing the residues at positions 243 and 264 to alanine. In oneembodiment, the Fc region is modified to decrease the ability of theantibody or fragment to mediate effector function and/or to increaseanti-inflammatory properties by modifying residues 243, 264, 267 and328.

In still another embodiment, the anti-LAG3 antibody (e.g., humanizedantibodies such as antagonist humanized antibodies) or antigen-bindingfragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) comprises a particular glycosylationpattern. For example, an aglycosylated antibody or fragment can be made(i.e., the antibody lacks glycosylation). The glycosylation pattern ofan antibody or fragment may be altered to, for example, increase theaffinity or avidity of the antibody or fragment for a LAG3 antigen. Suchmodifications can be accomplished by, for example, altering one or moreof the glycosylation sites within the antibody or fragment sequence. Forexample, one or more amino acid substitutions can be made that resultremoval of one or more of the variable region framework glycosylationsites to thereby eliminate glycosylation at that site. Suchaglycosylation may increase the affinity or avidity of the antibody orfragment for antigen. See, e.g., U.S. Pat. Nos. 5,714,350 and 6,350,861.

Anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments disclosed herein(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) include those produced in lower eukaryote hostcells, in particular fungal host cells such as yeast (e.g., Pichiapastoris) and filamentous fungi, that have been genetically engineeredto produce glycoproteins that have mammalian- or human-likeglycosylation patterns (See for example, Choi et al, (2003) Proc. Natl.Acad. Sci. 100: 5022-5027; Hamilton et al., (2003) Science 301:1244-1246; Hamilton et al., (2006) Science 313: 1441-1443). A particularadvantage of these genetically modified host cells over currently usedmammalian cell lines is the ability to control the glycosylation profileof glycoproteins that are produced in the cells such that compositionsof glycoproteins can be produced wherein a particular N-glycan structurepredominates (see, e.g., U.S. Pat. Nos. 7,029,872 and 7,449,308). Thesegenetically modified host cells have been used to produce antibodiesthat have predominantly particular N-glycan structures (See for example,Li et al., (2006) Nat. Biotechnol. 24: 210-215).

In particular embodiments, the anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) and antigen-bindingfragments thereof disclosed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) further includethose produced in lower eukaryotic host cells and which comprisefucosylated and non-fucosylated hybrid and complex N-glycans, includingbisected and multiantennary species, including but not limited toN-glycans such as GlcNAc₍₁₋₄₎Man₃GlcNAc₂;Gal₍₁₋₄₎GlcNAc₍₁₋₄₎Man₃GlcNAc₂; NANA₍₁₋₄₎Gal₍₁₋₄₎GlcNAc₍₁₋₄₎Man₃GlcNAc₂.

In particular embodiments, the anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) and antigen-bindingfragments thereof provided herein (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may compriseantibodies or fragments having at least one hybrid N-glycan selectedfrom the group consisting of GlcNAcMan₅GlcNAc₂; GalGlcNAcMan₅GlcNAc₂;and NANAGalGlcNAcMan₅GlcNAc₂. In particular aspects, the hybrid N-glycanis the predominant N-glycan species in the composition. In furtheraspects, the hybrid N-glycan is a particular N-glycan species thatcomprises about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%,or 100% of the hybrid N-glycans in the composition.

In particular embodiments, the anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) and antigen-bindingfragments thereof provided herein (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) compriseantibodies and fragments having at least one complex N-glycan selectedfrom the group consisting of GlcNAcMan₃GlcNAc₂; GalGlcNAcMan₃GlcNAc₂;NANAGalGlcNAcMan₃GlcNAc₂; GlcNAc₂Man₃GlcNAc₂; GalGlcNAc₂Man₃GlcNAc₂;Gal₂GlcNAc₂Man₃GlcNAc₂; NANAGal₂GlcNAc₂Man₃GlcNAc₂; andNANA₂Gal₂GlcNAc₂Man₃GlcNAc₂. In particular aspects, the complex N-glycanis the predominant N-glycan species in the composition. In furtheraspects, the complex N-glycan is a particular N-glycan species thatcomprises about 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%,or 100% of the complex N-glycans in the composition.

In particular embodiments, the anti-LAG3 antibody and antigen-bindingfragment N-glycan is fucosylated. In general, the fucose is in anα1,3-linkage with the GlcNAc at the reducing end of the N-glycan, anα1,6-linkage with the GlcNAc at the reducing end of the N-glycan, anα1,2-linkage with the Gal at the non-reducing end of the N-glycan, anα1,3-linkage with the GlcNac at the non-reducing end of the N-glycan, oran α1,4-linkage with a GlcNAc at the non-reducing end of the N-glycan.

Therefore, in particular aspects of the above the glycoproteincompositions, the glycoform is in an α1,3-linkage or α1,6-linkage fucoseto produce a glycoform selected from the group consisting ofMan₅GlcNAc₂(Fuc), GlcNAcMan₅GlcNAc₂(Fuc), Man₃GlcNAc₂(Fuc),GlcNAcMan₃GlcNAc₂(Fuc), GlcNAc₂Man₃GlcNAc₂(Fuc),GalGlcNAc₂Man₃GlcNAc₂(Fuc), Gal₂GlcNAc₂Man₃GlcNAc₂(Fuc),NANAGal₂GlcNAc₂Man₃GlcNAc₂(Fuc), and NANA₂Gal₂GlcNAc₂Man₃GlcNAc₂(Fuc);in an α1,3-linkage or α1,4-linkage fucose to produce a glycoformselected from the group consisting of GlcNAc(Fuc)Man₅GlcNAc₂,GlcNAc(Fuc)Man₃GlcNAc₂, GlcNAc₂(Fuc₁₋₂)Man₃GlcNAc₂,GalGlcNAc₂(Fuc₁₋₂)Man₃GlcNAc₂, Gal₂GlcNAc₂(Fuc₁₋₂)Man3GlcNAc₂,NANAGal₂GlcNAc₂(Fuc₁₋₂)Man₃GlcNAc₂, andNANA₂Gal₂GlcNAc₂(Fuc₁₋₂)Man₃GlcNAc₂; or in an aα1,2-linkage fucose toproduce a glycoform selected from the group consisting ofGal(Fuc)GlcNAc₂Man₃GlcNAc₂, Gal₂(Fuc₁₋₂)GlcNAc₂Man₃GlcNAc₂,NANAGal₂(Fuc₁₋₂)GlcNAc₂Man₃GlcNAc₂, andNANA₂Gal₂(Fuc₁₋₂)GlcNAc₂Man₃GlcNAc₂.

In further aspects, the anti-LAG3 antibodies (e.g., humanized antibodiessuch as antagonist humanized antibodies) or antigen-binding fragmentsthereof comprise high mannose N-glycans, including but not limited to,Man₈GlcNAc₂, Man₇GlcNAc₂, Man₆GlcNAc₂, Man₅GlcNAc₂, Man₄GlcNAc₂, orN-glycans that consist of the Man₃GlcNAc₂ N-glycan structure.

In further aspects of the above, the complex N-glycans further includefucosylated and non-fucosylated bisected and multiantennary species.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 or Ab9) comprises an immunoglobulin Fc domain that comprisesglycans that comprise sialic acid (e.g., N-Acetylneuraminic acid), e.g.,terminal α2,3-sialic acid or terminal α2,6-sialic acid. In an embodimentof the invention, the glycans on the Fc are 5, 10, 20, 50, 90% or moresialylated species. In an embodiment of the invention, the Fc comprisesthe mutations at positions 297, 264 and/or 243.

As used herein, the terms “N-glycan” and “glycoform” are usedinterchangeably and refer to an N-linked oligosaccharide, for example,one that is attached by an asparagine-N-acetylglucosamine linkage to anasparagine residue of a polypeptide. N-linked glycoproteins contain anN-acetylglucosamine residue linked to the amide nitrogen of anasparagine residue in the protein. The predominant sugars found onglycoproteins are glucose, galactose, mannose, fucose,N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc) and sialicacid (e.g., N-acetyl-neuraminic acid (NANA)). The processing of thesugar groups occurs co-translationally in the lumen of the ER andcontinues post-translationally in the Golgi apparatus for N-linkedglycoproteins.

N-glycans have a common pentasaccharide core of Man₃GlcNAc₂ (“Man”refers to mannose; “Glc” refers to glucose; and “NAc” refers toN-acetyl; GlcNAc refers to N-acetylglucosamine). Usually, N-glycanstructures are presented with the non-reducing end to the left and thereducing end to the right. The reducing end of the N-glycan is the endthat is attached to the Asn residue comprising the glycosylation site onthe protein. N-glycans differ with respect to the number of branches(antennae) comprising peripheral sugars (e.g., GlcNAc, galactose, fucoseand sialic acid) that are added to the Man₃GlcNAc₂ (“Man3”) corestructure which is also referred to as the “trimannose core”, the“pentasaccharide core” or the “paucimannose core”. N-glycans areclassified according to their branched constituents (e.g., high mannose,complex or hybrid). A “high mannose” type N-glycan has five or moremannose residues. A “complex” type N-glycan typically has at least oneGlcNAc attached to the 1,3 mannose arm and at least one GlcNAc attachedto the 1,6 mannose arm of a “trimannose” core. Complex N-glycans mayalso have galactose (“Gal”) or N-acetylgalactosamine (“GalNAc”) residuesthat are optionally modified with sialic acid or derivatives (e.g.,“NANA” or “NeuAc”, where “Neu” refers to neuraminic acid and “Ac” refersto acetyl). Complex N-glycans may also have intrachain substitutionscomprising “bisecting” GlcNAc and core fucose (“Fuc”). Complex N-glycansmay also have multiple antennae on the “trimannose core,” often referredto as “multiple antennary glycans.” A “hybrid” N-glycan has at least oneGlcNAc on the terminal of the 1,3 mannose arm of the trimannose core andzero or more mannoses on the 1,6 mannose arm of the trimannose core. Thevarious N-glycans are also referred to as “glycoforms.”

With respect to complex N-glycans, the terms “G-2”, “G-1”, “G0”, “G1”,“G2”, “A1”, and “A2” mean the following. “G-2” refers to an N-glycanstructure that can be characterized as Man₃GlcNAc₂; the term “G-1”refers to an N-glycan structure that can be characterized asGlcNAcMan₃GlcNAc₂; the term “G0” refers to an N-glycan structure thatcan be characterized as GlcNAc₂Man₃GlcNAc₂; the term “G1” refers to anN-glycan structure that can be characterized as GalGlcNAc₂Man₃GlcNAc₂;the term “G2” refers to an N-glycan structure that can be characterizedas Gal₂GlcNAc₂Man₃GlcNAc₂; the term “A1” refers to an N-glycan structurethat can be characterized as NANAGal₂GlcNAc₂Man₃GlcNAc₂; and, the term“A2” refers to an N-glycan structure that can be characterized asNANA₂Gal₂GlcNAc₂Man₃GlcNAc₂. Unless otherwise indicated, the terms G-2″,“G-1”, “G0”, “G1”, “G2”, “A1”, and “A2” refer to N-glycan species thatlack fucose attached to the GlcNAc residue at the reducing end of theN-glycan. When the term includes an “F”, the “F” indicates that theN-glycan species contains a fucose residue on the GlcNAc residue at thereducing end of the N-glycan. For example, G0F, G1F, G2F, A1F, and A2Fall indicate that the N-glycan further includes a fucose residueattached to the GlcNAc residue at the reducing end of the N-glycan.Lower eukaryotes such as yeast and filamentous fungi do not normallyproduce N-glycans that produce fucose.

With respect to multiantennary N-glycans, the term “multiantennaryN-glycan” refers to N-glycans that further comprise a GlcNAc residue onthe mannose residue comprising the non-reducing end of the 1,6 arm orthe 1,3 arm of the N-glycan or a GlcNAc residue on each of the mannoseresidues comprising the non-reducing end of the 1,6 arm and the 1,3 armof the N-glycan. Thus, multiantennary N-glycans can be characterized bythe formulas GlcNAc₍₂₋₄₎Man₃GlcNAc₂, Gal₍₁₋₄₎GlcNAc₍₂₋₄₎Man₃GlcNAc₂, orNANA₍₁₋₄₎Gal₍₁₋₄₎GlcNAc₍₂₋₄₎Man3GlcNAc₂. The term “1-4” refers to 1, 2,3, or 4 residues.

With respect to bisected N-glycans, the term “bisected N-glycan” refersto N-glycans in which a GlcNAc residue is linked to the mannose residueat the reducing end of the N-glycan. A bisected N-glycan can becharacterized by the formula GlcNAc₃Man₃GlcNAc₂ wherein each mannoseresidue is linked at its non-reducing end to a GlcNAc residue. Incontrast, when a multiantennary N-glycan is characterized asGlcNAc₃Man₃GlcNAc₂, the formula indicates that two GlcNAc residues arelinked to the mannose residue at the non-reducing end of one of the twoarms of the N-glycans and one GlcNAc residue is linked to the mannoseresidue at the non-reducing end of the other arm of the N-glycan.

Antibody Physical Properties

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may further contain one or moreglycosylation sites in either the light or heavy chain immunoglobulinvariable region. Such glycosylation sites may result in increasedimmunogenicity of the antibody or fragment or an alteration of the pK ofthe antibody due to altered antigen-binding (Marshall et al. (1972) AnnuRev Biochem 41:673-702; Gala and Morrison (2004) J Immunol 172:5489-94;Wallick et al (1988) J Exp Med 168:1099-109; Spiro (2002) Glycobiology12:43R-56R; Parekh et al (1985) Nature 316:452-7; Mimura et al. (2000)Mol Immunol 37:697-706). Glycosylation has been known to occur at motifscontaining an N-X-S/T sequence.

Each anti-LAG3 antibody (e.g., humanized antibodies such as antagonisthumanized antibodies) or antigen-binding fragment (e.g., 4A10, 19E8,11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/orAb9) will have a unique isoelectric point (pI). For example, someantibodies, such as Ab6, have a pI of about 6.3.

Each anti-LAG3 antibody (e.g., humanized antibodies such as antagonisthumanized antibodies) or antigen-binding fragment (e.g., 4A10, 19E8,11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/orAb9) will have a characteristic melting temperature, with a highermelting temperature indicating greater overall stability in vivo(Krishnamurthy R and Manning M C (2002) Curr Pharm Biotechnol 3:361-71).In general, the T_(M1) (the temperature of initial unfolding) may begreater than 60° C., greater than 65° C., or greater than 70° C. Themelting point of an antibody or fragment can be measured usingdifferential scanning calorimetry (Chen et al (2003) Pharm Res20:1952-60; Ghirlando et al (1999) Immunol Lett 68:47-52) or circulardichroism (Murray et al. (2002) J. Chromatogr Sci 40:343-9).

In a further embodiment, anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) and antigen-bindingfragments thereof (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) are selected that do notdegrade rapidly. Degradation of an antibody or fragment can be measuredusing capillary electrophoresis (CE) and MALDI-MS (Alexander A J andHughes D E (1995) Anal Chem 67:3626-32).

In a further embodiment, anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) and antigen-bindingfragments thereof are selected that have minimal aggregation effects,which can lead to the triggering of an unwanted immune response and/oraltered or unfavorable pharmacokinetic properties. Generally, antibodiesand fragments are acceptable with aggregation of 25% or less, 20% orless, 15% or less, 10% or less, or 5% or less. Aggregation can bemeasured by several techniques, including size-exclusion column (SEC),high performance liquid chromatography (HPLC), and light scattering.

Antibody Conjugates

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be conjugated to a chemicalmoiety. Such conjugated antibodies and fragments are part of the presentinvention. The chemical moiety may be, inter alia, a polymer, aradionuclide or a cytotoxic factor. In particular embodiments, thechemical moiety is a polymer which increases the half-life of theantibody or fragment in the body of a subject. Suitable polymersinclude, but are not limited to, hydrophilic polymers which include butare not limited to polyethylene glycol (PEG) (e.g., PEG with a molecularweight of 2 kDa, 5 kDa, 10 kDa, 12 kDa, 20 kDa, 30 kDa or 40 kDa),dextran and monomethoxypolyethylene glycol (mPEG). Lee, et al., (1999)(Bioconj. Chem. 10:973-981) discloses PEG conjugated single-chainantibodies. Wen, et al., (2001) (Bioconj. Chem. 12:545-553) discloseconjugating antibodies with PEG which is attached to a radiometalchelator (diethylenetriaminpentaacetic acid (DTPA)).

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be conjugated with labels suchas ⁹⁹Tc, ⁹⁰Y, ¹¹¹In, ³²P, ¹⁴C, ¹²⁵I, ³H, ¹³¹I, ¹¹C, ¹⁵O, ¹³N, ¹⁸F, ³⁵S,⁵¹Cr, ⁵⁷To, ²²⁶Ra, ⁶⁰Co, ⁵⁹Fe, ⁵⁷Se, ¹⁵²Eu, ⁶⁷CU, ²¹⁷Ci, ²¹¹At, ²¹²Pb,⁴⁷Sc, ¹⁰⁹Pd, ²³⁴Th, and ⁴⁰K, ¹⁵⁷Gd, ⁵⁵Mn, ⁵²Tr, and ⁵⁶Fe.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments disclosed herein(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) may also be PEGylated, for example to increase itsbiological (e.g., serum) half-life. To PEGylate an antibody or fragment,the antibody or fragment, typically is reacted with a reactive form ofpolyethylene glycol (PEG), such as a reactive ester or aldehydederivative of PEG, under conditions in which one or more PEG groupsbecome attached to the antibody or antibody fragment. In particularembodiments, the PEGylation is carried out via an acylation reaction oran alkylation reaction with a reactive PEG molecule (or an analogousreactive water-soluble polymer). As used herein, the term “polyethyleneglycol” is intended to encompass any of the forms of PEG that have beenused to derivatize other proteins, such as mono (C1-C10) alkoxy- oraryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certainembodiments, the antibody or fragment to be PEGylated is anaglycosylated antibody or fragment. Methods for PEGylating proteins areknown in the art and can be applied to the antibodies of the invention.See, e.g., EP 0 154 316 and EP 0 401 384.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments disclosed herein(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) may also be conjugated with fluorescent orchemiluminescent labels, including fluorophores such as rare earthchelates, fluorescein and its derivatives, rhodamine and itsderivatives, isothiocyanate, phycoerythrin, phycocyanin,allophycocyanin, o-phthaladehyde, fluorescamine, ¹⁵²Eu, dansyl,umbelliferone, luciferin, luminal label, isoluminal label, an aromaticacridinium ester label, an imidazole label, an acridimium salt label, anoxalate ester label, an aequorin label, 2,3-dihydrophthalazinediones,biotin/avidin, spin labels and stable free radicals.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) may also be conjugated to a cytotoxic factor such asdiptheria toxin, Pseudomonas aeruginosa exotoxin A chain, ricin A chain,abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteinsand compounds (e.g., fatty acids), dianthin proteins, Phytoiaccaamericana proteins PAPI, PAPII, and PAP-S, momordica charantiainhibitor, curcin, crotin, Saponaria officinalis inhibitor, mitogellin,restrictocin, phenomycin, and enomycin.

Any method known in the art for conjugating the anti-LAG3 antibodies(e.g., humanized antibodies such as antagonist humanized antibodies) andantigen-binding fragments thereof (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to the variousmoieties may be employed, including those methods described by Hunter,et al., (1962) Nature 144:945; David, et al., (1974) Biochemistry13:1014; Pain, et al., (1981) J. Immunol. Meth. 40:219; and Nygren, J.,(1982) Histochem. and Cytochem. 30:407. Methods for conjugatingantibodies and fragments are conventional and very well known in theart.

Therapeutic Uses of Anti-LAG3 Antibodies

Further provided are methods for treating or preventing cancer insubjects, such as human subjects, in need of such treatment byadministering an effective amount of the anti-LAG3 antibodies orantigen-binding fragments thereof of the present invention which aredisclosed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) which may be effective for suchtreatment or prevention. In an embodiment of the invention, such asubject suffers from and is treated for cancer, e.g., a solid tumorwhich includes, in addition to the tumor cells, tumor infiltratinglymphocytes (TILs), such as T-cells, expressing LAG3, e.g.,osteosarcoma, rhabdomyosarcoma, neuroblastoma, kidney cancer, leukemia,renal transitional cell cancer, bladder cancer, Wilm's cancer, ovariancancer, pancreatic cancer, breast cancer (e.g., characterized by amutation in BRCA1 and/or BRCA2), prostate cancer, bone cancer, lungcancer (e.g., non-small cell lung cancer), gastric cancer, colorectalcancer, cervical cancer, synovial sarcoma, head and neck cancer,squamous cell carcinoma, multiple myeloma, renal cell cancer,retinoblastoma, hepatoblastoma, hepatocellular carcinoma, melanoma,rhabdoid tumor of the kidney, Ewing's sarcoma, chondrosarcoma, braincancer, glioblastoma, meningioma, pituitary adenoma, vestibularschwannoma, a primitive neuroectodermal tumor, medulloblastoma,astrocytoma, anaplastic astrocytoma, oligodendroglioma, ependymoma,choroid plexus papilloma, polycythemia vera, thrombocythemia, idiopathicmyelfibrosis, soft tissue sarcoma, thyroid cancer, endometrial cancer,carcinoid cancer or liver cancer, breast cancer or gastric cancer. In anembodiment of the invention, the cancer is metastatic cancer, e.g., ofthe varieties described above.

The present invention also provides methods for treating or preventingan infectious disease in a subject by administering an effective amountof anti-LAG3 antibodies or antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to the subject which may be effective forsuch treatment or prevention. In an embodiment of the invention, theinfectious disease is viral infection. In an embodiment of theinvention, the infectious disease is bacterial infection. In anembodiment of the invention, the infectious disease is parasiticinfection. In an embodiment of the invention, the infectious disease isfungal infection.

The present invention includes methods of treating any of the cancers orinfectious diseases discussed herein by administering a therapeuticallyeffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) optionally in association with any of thefurther chemotherapeutic agents or therapeutic procedures discussedherein as well as compositions including such an antibody or fragment inassociation with such a further chemotherapeutic agent.

In an embodiment of the invention, the viral infection is infection witha virus selected from the group consisting of human immunodeficiencyvirus (HIV), ebola virus, hepatitis virus (A, B, or C), herpes virus(e.g., VZV, HSV-I, HAV-6, HSV-II, and CMV, Epstein Barr virus),adenovirus, influenza virus, flaviviruses, echovirus, rhinovirus,coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus,rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus,HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus,rabies virus, JC virus or arboviral encephalitis virus.

In an embodiment of the invention, the bacterial infection is infectionwith a bacteria selected from the group consisting of Chlamydia,rickettsial bacteria, mycobacteria, staphylococci, streptococci,pneumonococci, meningococci and gonococci, klebsiella, proteus,serratia, pseudomonas, Legionella, Corynebacterium diphtherias,Salmonella, bacilli, Vibrio cholerae, Clostridium tetan, Clostridiumbotulinum, Bacillus anthricis, Yersinia pestis, Mycobacterium leprae,Mycobacterium lepromatosis, and Borriella.

In an embodiment of the invention, the fungal infection is infectionwith a fungus selected from the group consisting of Candida (albicans,krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans,Aspergillus (fumigatus, niger, etc.), Genus Mucorales (mucor, absidia,rhizopus), Sporothrix schenkii, Blastomyces dermatitidis,Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasmacapsulatum.

In an embodiment of the invention, the parasitic infection is infectionwith a parasite selected from the group consisting of Entamoebahistolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba, GiardiaZambia, Cryptosporidium, Pneumocystis carinii, Plasmodium vivax, Babesiamicroti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani,Toxoplasma gondii, Nippostrongylus brasiliensis.

In addition, the present invention provides a method for preventing orinhibiting LAG3 binding to MEW class II, enhancing antigen-specificT-cell activation or stimulating T-cell production of interleukin-2 in asubject (e.g., human), for example, wherein the subject suffers fromcancer or infectious disease (e.g., as discussed herein) comprisingadministering an effective amount of anti-LAG3 antibody orantigen-binding fragment thereof (e.g., 4A10, 19E8, 11C9, 22D2, Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9, to the subject,optionally, in association with a further chemotherapeutic agent, e.g.,pembrolizumab or nivolumab.

The scope of the present invention provides uses of the anti-LAG3antibodies or antigen-binding fragments thereof disclosed herein (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) in the manufacture of a medicament for treating canceror infectious disease in a subject.

The present invention includes methods for treating or preventingosteosarcoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingrhabdomyosarcoma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingneuroblastoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing kidneycancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingleukemia comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing renaltransitional cell cancer comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingbladder cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing Wilm'scancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingovarian cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingpancreatic cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing breastcancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof. In anembodiment of the invention, the method for treating or preventingbreast cancer comprises administering an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention in association with an anthracycline (e.g., doxorubicin and/orepirubicin) and/or a taxane (e.g., paclitaxel and/or docetaxel).Optionally, an anthracycline and taxane is in association with5-fluorouracil (5-FU), cyclophosphamide, and carboplatin. In anembodiment of the invention, wherein the breast cancer is HER2 positive,the anti-LAG3 antibody or fragment is administered in association withtrastuzumab, optionally with a taxane and/or pertuzumab.

The present invention includes methods for treating or preventingprostate cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing bonecancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing lungcancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof. In anembodiment of the invention, the method for treating or preventing lungcancer comprises administering an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention inassociation with bevacizumab and/or cetuximab.

The present invention includes methods for treating or preventingnon-small cell lung cancer comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof. In an embodiment of the invention, the method for treating orpreventing non-small cell lung cancer comprises administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention in association with cisplatin,carboplatin, paclitaxel, albumin-bound paclitaxel, docetaxel,gemcitabine, vinorelbine, irinotecan, etoposide, vinblastine, and/orpemetrexed.

The present invention includes methods for treating or preventinggastric cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingcolorectal cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof. Inan embodiment of the invention, the method for treating or preventingcolorectal cancer comprises administering an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention in association with 5-Fluorouracil (5-FU), capecitabine,irinotecan and/or oxaliplatin (e.g., FOLFOX, FOLFIRI, FOLFOXIRI orCapeOx).

The present invention includes methods for treating or preventingcervical cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingsynovial sarcoma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing headand neck cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingsquamous cell carcinoma comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingmultiple myeloma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing renalcell cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingretinoblastoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventinghepatoblastoma comprising administering (optionally in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventinghepatocellular carcinoma comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingmelanoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingrhabdoid tumor of the kidney comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingEwing's sarcoma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingchondrosarcoma comprising administering (optionally in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing braincancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingglioblastoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof. In anembodiment of the invention, the method for treating or preventingglioblastoma multiforme comprises administering an effective amount ofan anti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention in association with temozolomide.

The present invention includes methods for treating or preventingmeningioma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingpituitary adenoma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingvestibular schwannoma comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventing aprimitive neuroectodermal tumor comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingmedulloblastoma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingastrocytoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventinganaplastic astrocytoma comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof. In an embodiment of the invention, the method for treating orpreventing refractory anaplastic astrocytoma comprises administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention in association with temozolomide.

The present invention includes methods for treating or preventingoligodendroglioma comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingependymoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingchoroid plexus papilloma comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventingpolycythemia vera comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingthrombocythemia comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingidiopathic myelfibrosis comprising administering (optionally, inassociation with pembrolizumab or nivolumab) an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof.

The present invention includes methods for treating or preventing softtissue sarcoma comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingthyroid cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingendometrial cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventingcarcinoid cancer comprising administering (optionally, in associationwith pembrolizumab or nivolumab) an effective amount of an anti-LAG3antibody or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing livercancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing breastcancer (e.g., characterized by a mutation in BRCA1 and/or BRCA2)comprising administering (optionally, in association with pembrolizumabor nivolumab) an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventinggastric cancer comprising administering (optionally, in association withpembrolizumab or nivolumab) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with human immunodeficiency virus (HIV) in a subjectcomprising administering) an effective amount of an anti-LAG3 antibodyor antigen-binding fragment thereof of the present invention (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) to a subject, such as a human, in need thereof.Optionally, the subject is administered an anti-viral therapeutic agentsuch as a protease inhibitor, a nucleoside/nucleotide reversetranscriptase inhibitor, a non-nucleoside reverse transcriptaseinhibitors, an entry inhibitor, a fusion inhibitor or an integraseinhibitors.

The present invention includes methods for treating or preventing aninfection with Bundibugyo virus (BDBV), Sudan virus (SUDV), Tai Forestvirus (TAFV) and/or ebola virus in a subject comprising administering)an effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent, such as one or moreantibodies that specifically bind to the BDBV, SUDV, TAFV or ebola virusor a nucleoside RNA polymerase inhibitor; or a vaccine.

The present invention includes methods for treating or preventing aninfection with hepatitis A virus in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with hepatitis B virus in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with hepatitis C virus in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent such as interferon and/orribavirin.

The present invention includes methods for treating or preventing aninfection with herpes virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with vesicular stomatitis virus in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with herpes simplex virus-I in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with HAV-6 virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with herpes simplex virus-II in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with cytomegalovirus (CMV) in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with epstein Barr virus in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with adenovirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with influenza virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with flavivirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with echovirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with rhinovirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with coxsackie virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with coronavirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with respiratory syncytial virus in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with mumps virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with rotavirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with measles virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with rubella virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with parvovirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with vaccinia virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with human T-lymphotropic virus (HTLV) in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with dengue virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with papillomavirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with molluscum virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with poliovirus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with rabies virus in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with John Cunningham virus (JC virus) in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with arboviral encephalitis virus in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-viral therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Chlamydia trachomatis in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with rickettsia bacteria in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with mycobacteria in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with staphylococci in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with streptococci in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with pneumonococci in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with meningococci in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with gonococci in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with klebsiella in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with proteus (e.g., P. vulgaris, P. mirabilis, or P. penneri)in a subject comprising administering an effective amount of ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof. Optionally, the subject is administered an anti-bacterialantibiotic.

The present invention includes methods for treating or preventing aninfection with serratia in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with pseudomonas in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with legionella in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Corynebacterium diphtheriae in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Salmonella (e.g., Salmonella bongori or Salmonellaenterica) in a subject comprising administering an effective amount ofan anti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject, such as a human, in needthereof. Optionally, the subject is administered an anti-bacterialantibiotic.

The present invention includes methods for treating or preventing aninfection with bacilli in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Vibrio cholerae in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Clostridium tetani in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Clostridium botulinum in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Bacillus anthracis in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Yersinia pestis in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Leptospira in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Borrelia in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-bacterial antibiotic.

The present invention includes methods for treating or preventing aninfection with Candida albicans in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Candida krusei in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Candida glabrata in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Candida tropicalis in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Cryptococcus neoformans in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Aspergillus fumigatus in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Aspergillus niger in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Mucorales mucor in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Mucorales absidia in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Mucorales rhizopus in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Sporothrix schenkii in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof. Optionally, the subject isadministered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Blastomyces dermatitidis in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Paracoccidioides brasiliensis in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Coccidioides immitis in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Histoplasma capsulatum in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof. Optionally,the subject is administered an anti-fungal therapeutic agent.

The present invention includes methods for treating or preventing aninfection with Entamoeba histolytica in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Balantidium coli in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Naegleria fowleri in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Acanthamoeba sp. in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Giardia Zambia in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Cryptosporidium sp. in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Pneumocystis carinii in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Plasmodium vivax in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Babesia microti in a subject comprising administering aneffective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Trypanosoma brucei in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Trypanosoma cruzi in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Leishmania donovani in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Toxoplasma gondii in a subject comprising administeringan effective amount of an anti-LAG3 antibody or antigen-binding fragmentthereof of the present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) to a subject,such as a human, in need thereof.

The present invention includes methods for treating or preventing aninfection with Nippostrongylus brasiliensis in a subject comprisingadministering an effective amount of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) to a subject, such as a human, in need thereof.

A “subject” is a mammal such as, for example, a human, dog, cat, horse,cow, mouse, rat, monkey (e.g., cynomolgous monkey, e.g., Macacafascicularis or Macaca mulatta) or rabbit.

In particular embodiments, the anti-LAG3 antibodies (e.g., humanizedantibodies such as antagonist humanized antibodies) or antigen-bindingfragments thereof of the present invention which are disclosed herein(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) may be used alone, or in association with other,further therapeutic agents and/or therapeutic procedures, for treatingor preventing any disease such as cancer, e.g., as discussed herein, ina subject in need of such treatment or prevention. Compositions or kits,e.g., pharmaceutical compositions comprising a pharmaceuticallyacceptable carrier, comprising such antibodies and fragments inassociation with further therapeutic agents are also part of the presentinvention.

In particular embodiments, the anti-LAG3 antibodies (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragments thereof of the present invention (e.g., 4A10, 19E8, 11C9and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9)may be used in association with an anti-cancer therapeutic agent orimmunomodulatory drug such as an immunomodulatory receptor inhibitor,e.g., an antibody or antigen-binding fragment thereof that specificallybinds to the receptor.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibodies such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with one or more of an inhibitors (e.g., asmall organic molecule or an antibody or antigen-binding fragmentthereof) such as: an MTOR (mammalian target of rapamycin) inhibitor, acytotoxic agent, a platinum agent a BRAF inhibitor, a CDK4/6 inhibitoran EGFR inhibitor, a VEGF inhibitor, a microtubule stabilizer, a taxane,a CD20 inhibitor, a CD52 inhibitor, a CD30 inhibitor, a RANK (Receptoractivator of nuclear factor kappa-B) inhibitor, a RANKL (Receptoractivator of nuclear factor kappa-B ligand) inhibitor, an ERK inhibitor,a MAP Kinase inhibitor, an AKT inhibitor, a MEK inhibitor, a PI3Kinhibitor, a HER1 inhibitor, a HER2 inhibitor, a HER3 inhibitor, a HER4inhibitor, a Bcl2 inhibitor, a CD22 inhibitor, a CD79b inhibitor, anErbB2 inhibitor, or a farnesyl protein transferase inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with one or more of: anti-PD1 (e.g.,pembrolizumab, nivolumab, CT-011), anti-PDL1, anti-CTLA4, anti-TIM3,anti-CS1, (e.g., elotuzumab), anti-KIR2DL1/2/3 (e.g., lirilumab),anti-CD27, anti-CD137 (e.g., urelumab), anti-GITR (e.g., TRX518),anti-PD-L1 (e.g., BMS-936559, MSB0010718C or MPDL3280A), anti-PD-L2,anti-ILT1, anti-ILT2, anti-ILT3, anti-ILT4, anti-ILT5, anti-ILT6,anti-ILT7, anti-ILT8, anti-CD40, anti-OX40, anti-CD137, anti-KIR2DL1,anti-KIR2DL2/3, anti-KIR2DL4, anti-KIR2DL5A, anti-KIR2DL5B,anti-KIR3DL1, anti-KIR3DL2, anti-KIR3DL3, anti-NKG2A, anti-NKG2C,anti-NKG2E, or any small organic molecule inhibitor of such targets;IL-10, anti-IL10, anti-TSLP (thymic stromal lymphopoietin) or PEGylatedIL-10.

In an embodiment of the invention, the molecular weight of thepolyethylene glycol (PEG) moiety, on a PEGylated IL-10 molecule, isabout 12,000 daltons or about 20,000 daltons. In an embodiment of theinvention, PEGylated IL-10 (e.g., PEGylated human IL-10) comprises oneor more polyethylene glycol molecules covalently attached via a linker(e.g., C₂₋₁₂ alkyl such as —CH₂CH₂CH₂—) to a single amino acid residueof a single subunit of IL-10, wherein said amino acid residue is thealpha amino group of the N-terminal amino acid residue or the epsilonamino group of a lysine residue. In an embodiment of the inventionPEGylated IL-10 is: (PEG)_(b)-L-NH-IL-10; wherein b is 1-9 and L is aC₂₋₁₂ alkyl linker moiety covalently attached to a nitrogen (N) of thesingle amino acid residue of the IL-10. In an embodiment of theinvention, the IL-10 of PEGylated IL-10 has the formula:[X—O(CH₂CH₂O)_(n)]_(b)-L-NH-IL-10, wherein X is H or C₁₋₄ alkyl; n is 20to 2300; b is 1 to 9; and L is a C₁₋₁₁ alkyl linker moiety which iscovalently attached to the nitrogen (N) of the alpha amino group at theamino terminus of one IL-10 subunit; provided that when b is greaterthan 1, the total of n does not exceed 2300. See U.S. Pat. No.7,052,686.

In an embodiment of the invention, the anti-IL-10 antibody orantigen-binding fragment thereof (e.g., humanized antibody) comprisesthe CDRs set forth below:

CDR-L1: (SEQ ID NO: 465) KTSQNIFENLA CDR-L2: (SEQ ID NO: 466) NASPLQACDR-L3: (SEQ ID NO: 467) HQYYSGYT CDR-H1: (SEQ ID NO: 468) GFTFSDYHMACDR-H2: (SEQ ID NO: 469) SITLDATYTYYRDSVRG CDR-H3: (SEQ ID NO: 470)HRGFSVWLDY(See U.S. Pat. No. 7,662,379)

In an embodiment of the invention, the anti-TSLP antibody orantigen-binding fragment thereof (e.g., humanized antibody) comprisesthe CDRs set forth below:

CDR-H1: (SEQ ID NO: 428) GYIFTDYAMH; CDR-H2: (SEQ ID NO: 429)TFIPLLDTSDYNQNFK; CDR-H3: (SEQ ID NO: 430) MGVTHSYVMDA; CDR-L1:(SEQ ID NO: 431) RASQPISISVH; CDR-L2: (SEQ ID NO: 432) FASQSIS; CDR-L3:(SEQ ID NO: 433) QQTFSLPYT;(see WO2008/76321)

In an embodiment of the invention, the anti-CD27 antibody orantigen-binding fragment thereof (e.g., humanized antibody) comprisesthe CDRs set forth below:

CDR-H1: (SEQ ID NO: 420) GFIIKATYMH; CDR-H2: (SEQ ID NO: 421)RIDPANGETKYDPKFQV; CDR-H3: (SEQ ID NO: 422) YAWYFDV; CDR-L1:(SEQ ID NO: 423) RASENIYSFLA; CDR-L2: (SEQ ID NO: 424) HAKTLAE; CDR-L3:(SEQ ID NO: 425) QHYYGSPLT;(See WO2012/04367).

Thus, the present invention includes compositions comprising ananti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) in association with pembrolizumab; aswell as methods for treating or preventing cancer in a subjectcomprising administering an effective amount of the anti-LAG3 antibodyor antigen-binding fragment thereof in association with pembrolizumab(e.g., pembrolizumab dosed at 200 mg once every three weeks) to thesubject. Optionally, the subject is also administered in associationwith a another further therapeutic agent.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a pembrolizumab antibody whichcomprises an immunoglobulin heavy chain (or CDR-H1, CDR-H2 and CDR-H3thereof) comprising the amino acid sequence:

(SEQ ID NO: 41) QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYREDMGEDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVESCSVMHEALHNHYTQKSLSLSLGK;and an immunoglobulin light chain (or CDR-L1, CDR-L2 and CDR-L3 thereof)comprising the amino acid sequence:

(SEQ ID NO: 42) EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLTYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTEGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNEYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an antibody comprising animmunoglobulin heavy chain (or CDR-H1, CDR-H2 and CDR-H3 thereof)comprising the amino acid sequence:

(SEO ID NO: 43) QVQLVESGGGVVQPGRSLRLDCKASGITESNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK;and an immunoglobulin light chain (or CDR-L1, CDR-L2 and CDR-L3 thereof)comprising the amino acid sequence:

(SEQ ID NO: 44) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with any one or more of: 13-cis-retinoicacid, 3-[5-(methylsulfonylpiperadinemethyl)-indolyl]-quinolone,4-hydroxytamoxifen, 5-deooxyuridine, 5′-deoxy-5-fluorouridine,5-fluorouracil, 6-mecaptopurine, 7-hydroxystaurosporine, A-443654,abirateroneacetate, abraxane, ABT-578, acolbifene, ADS-100380,aflibercept, ALT-110, altretamine, amifostine, aminoglutethimide,amrubicin, amsacrine, anagrelide, anastrozole, angiostatin, AP-23573,ARQ-197, arzoxifene, AS-252424, AS-605240, asparaginase, ATI3387,AT-9263, atrasentan, axitinib, AZD1152, Bacillus Calmette-Guerin (BCG)vaccine, batabulin, BC-210, besodutox, bevacizumab, BGJ398,bicalutamide, Bio111, BIO140, BKM120, bleomycin, BMS-214662, BMS-247550,BMS-275291, BMS-310705, bortezimib, buserelin, busulfan, calcitriol,camptothecin, canertinib, capecitabine, carboplatin, carmustine, CC8490,CEA (recombinant vaccinia-carcinoembryonic antigen vaccine), cediranib,CG-1521, CG-781, chlamydocin, chlorambucil, chlorotoxin, cilengitide,cimitidine, cisplatin, cladribine, clodronate, cobimetnib, COL-3,CP-724714, cyclophosphamide, cyproterone, cyproteroneacetate,cytarabine, cytosinearabinoside, dabrafenib, dacarbazine, dacinostat,dactinomycin, dalotuzumab, danusertib, dasatanib, daunorubicin,decatanib, deguelin, denileukin, deoxycoformycin, depsipeptide,diarylpropionitrile, diethylstilbestrol, diftitox, DNE03, docetaxel,dovitinib, doxorubicin, droloxifene, edotecarin, yttrium-90labeled-edotreotide, edotreotide, EKB-569, EMD121974, encorafenib,endostatin, enzalutamide, enzastaurin, epirubicin, epithilone B,ERA-923, erbitux, erlotinib, estradiol, estramustine, etoposide,everolimus, exemestane, ficlatuzumab, finasteride, flavopiridol,floxuridine, fludarabine, fludrocortisone, fluoxymesterone, flutamide,FOLFOX regimen, fulvestrant, galeterone, ganetespib, gefitinib,gemcitabine, gimatecan, glucopyranosyl lipid A, goserelin, goserelinacetate, gossypol, GSK461364, GSK690693, EMIR-3339,hydroxyprogesteronecaproate, hydroxyurea, IC87114, idarubicin,idoxyfene, ifosfamide, IM862, imatinib, IMC-1C11, imiquimod, INC280,INCB24360, INO1001, interferon, interleukin-2, interleukin-12,ipilimumab, irinotecan, JNJ-16241199, ketoconazole, KRX-0402, lapatinib,lasofoxifene, LEE011, letrozole, leucovorin, leuprolide, leuprolideacetate, levamisole, liposome entrapped paclitaxel, lomustine,lonafarnib, lucanthone, LY292223, LY292696, LY293646, LY293684,LY294002, LY317615, LY3009120, marimastat, mechlorethamine,medroxyprogesteroneacetate, megestrolacetate, MEK162, melphalan,mercaptopurine, mesna, methotrexate, mithramycin, mitomycin, mitotane,mitoxantrone, a suspension of heat killed Mycobacterium obuense,tozasertib, MLN8054, natitoclax, neovastat, Neratinib, neuradiab,nilotinib, nilutimide, nolatrexed, NVP-BEZ235, oblimersen, octreotide,ofatumumab, oregovomab, ornatuzumab, orteronel, oxaliplatin, paclitaxel,palbociclib, pamidronate, panitumumab, pazopanib, PD0325901, PD184352,PEG-interferon, pemetrexed, pentostatin, perifosine,phenylalaninemustard, PI-103, pictilisib, PIK-75, pipendoxifene,PKI-166, plicamycin, poly-ICLC, porfimer, prednisone, procarbazine,progestins, PSK protein bound polysaccharide (derived from Basidiomycetecoriolus versicolor), PLX8394, PX-866, R-763, raloxifene, raltitrexed,razoxin, ridaforolimus, rituximab, romidepsin, RTA744, rubitecan,scriptaid, Sdx102, seliciclib, selumetinib, semaxanib, SF1126,sirolimus, SN36093, sorafenib, spironolactone, squalamine, SR13668,streptozocin, SU6668, suberoylanalide hydroxamic acid, sunitinib,synthetic estrogen, talampanel, talimogene laherparepvec, tamoxifen,temozolomide, temsirolimus, teniposide, tesmilifene, testosterone,tetrandrine, TGX-221, thalidomide, 6-thioguanine, thiotepa, ticilimumab,tipifarnib, tivozanib, TKI-258, TLK286, TNFα (tumor necrosis factoralpha), topotecan, toremifene citrate, trabectedin, trametinib,trastuzumab, tretinoin, trichostatin A, triciribinephosphatemonohydrate, triptorelin pamoate, TSE-424, uracil mustard, valproicacid, valrubicin, vandetanib, vatalanib, VEGF trap, vemurafenib,vinblastine, vincristine, vindesine, vinorelbine, vitaxin, vitespan,vorinostat, VX-745, wortmannin, Xr311, Z-100 hot water extract ofBacillus tuberculosis, zanolimumab, ZK186619, ZK-304709, ZM336372 orZSTK474.

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with one or more antiemetics including,but not limited to: casopitant (GlaxoSmithKline), Netupitant(MGI-Helsinn) and other NK-1 receptor antagonists, palonosetron (sold asAloxi by MGI Pharma), aprepitant (sold as Emend by Merck and Co.;Rahway, N.J.), diphenhydramine (sold as Benadryl® by Pfizer; New York,N.Y.), hydroxyzine (sold as Atarax® by Pfizer; New York, N.Y.),metoclopramide (sold as Reglan® by AH Robins Co.; Richmond, Va.),lorazepam (sold as Ativan® by Wyeth; Madison, N.J.), alprazolam (sold asXanax® by Pfizer; New York, N.Y.), haloperidol (sold as Haldol® byOrtho-McNeil; Raritan, N.J.), droperidol (Inapsine®), dronabinol (soldas Marinol® by Solvay Pharmaceuticals, Inc.; Marietta, Ga.),dexamethasone (sold as Decadron® by Merck and Co.; Rahway, N.J.),methylprednisolone (sold as Medrol® by Pfizer; New York, N.Y.),prochlorperazine (sold as Compazine® by Glaxosmithkline; ResearchTriangle Park, N.C.), granisetron (sold as Kytril® by Hoffmann-La RocheInc.; Nutley, N.J.), ondansetron (sold as Zofran® by by Glaxosmithkline;Research Triangle Park, N.C.), dolasetron (sold as Anzemet® bySanofi-Aventis; New York, N.Y.), tropisetron (sold as Navoban® byNovartis; East Hanover, N.J.).

Other side effects of cancer treatment include red and white blood celldeficiency. Accordingly, in an embodiment of the invention, an anti-LAG3antibody (e.g., humanized antibody such as antagonist humanizedantibodies) or antigen-binding fragment thereof of the present invention(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9) is in association with an agent which treats orprevents such a deficiency, such as, e.g., filgrastim, PEG-filgrastim,erythropoietin, epoetin alfa or darbepoetin alfa.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a vaccine. In an embodiment of theinvention, the vaccine is an anti-cancer vaccine, a peptide vaccine or aDNA vaccine. For example, in an embodiment of the invention, the vaccineis a tumor cell (e.g., an irradiated tumor cell) or a dendritic cell(e.g., a dendritic cell pulsed with a tumor peptide).

In an embodiment of the invention, an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is administered in association with a therapeutic procedure.A therapeutic procedure is one or more steps carried out by a physicianor clinician in treating a subject which is intended to alleviate one ormore symptoms (e.g., of cancer and/or infectious disease) in the treatedsubject, whether by inducing the regression or elimination of suchsymptoms or by inhibiting the progression of such symptom(s), e.g.,cancer symptoms such as tumor growth or metastasis, by any clinicallymeasurable degree.

In an embodiment of the invention, a therapeutic procedure isanti-cancer radiation therapy. For example, in an embodiment of theinvention, the radiation therapy is external beam therapy (EBT): amethod for delivering a beam of high-energy X-rays to the location ofthe tumor. The beam is generated outside the patient (e.g., by a linearaccelerator) and is targeted at the tumor site. These X-rays can destroythe cancer cells and careful treatment planning allows the surroundingnormal tissues to be spared. No radioactive sources are placed insidethe patient's body. In an embodiment of the invention, the radiationtherapy is proton beam therapy: a type of conformal therapy thatbombards the diseased tissue with protons instead of X-rays. In anembodiment of the invention, the radiation therapy is conformal externalbeam radiation therapy: a procedure that uses advanced technology totailor the radiation therapy to an individual's body structures.

In an embodiment of the invention, the radiation therapy isbrachytherapy: the temporary placement of radioactive materials withinthe body, usually employed to give an extra dose—or boost—of radiationto an area.

In an embodiment of the invention, a surgical procedure administered inassociation with an anti-LAG3 antibody (e.g., humanized antibody such asantagonist humanized antibodies) or antigen-binding fragment thereof ofthe present invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) is surgical tumorectomy.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an MTOR (mammalian target ofrapamycin) inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a cytotoxic agent.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a platinum agent.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an EGFR inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a VEGF inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a microtubule stabilizer.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a taxane a CD20 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a CD52 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a CD30 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a RANK (Receptor activator of nuclearfactor kappa-B) inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a RANKL (Receptor activator ofnuclear factor kappa-B ligand) inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a BRAF inhibitor, e.g., for treatmentof melanoma.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an a CDK4/6 inhibitor, e.g., fortreatment of melanoma.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an ERK inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a MAP Kinase inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an AKT inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a MEK inhibitor, e.g., for treatmentof melanoma.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a PI3K inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a HER1 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a HER2 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a HER3 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a HER4 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a Bcl2 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a CD22 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a CD79b inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an ErbB2 inhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a farnesyl protein transferaseinhibitor.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-PD1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with nivolumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CT-011.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-PDL1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-CTLA4.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-TIM3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-CS1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with elotuzumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL1/2/3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lirilumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an anti-CD137 antibody orantigen-binding fragment thereof, e.g., an agonist anti-CD137 antibodyor fragment.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with urelumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-GITR.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with TRX518.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment (e.g., humanized antibody such as antagonisthumanized antibodies) of the present invention (e.g., 4A10, 19E8, 11C9and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) isin association with anti-PD-L1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BMS-936559.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with MSB0010718C.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with MPDL3280A.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-PD-L2.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT2.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT4.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT5.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT6.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT7.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-ILT8.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-CD40.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-OX40.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-CD137.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such asantagonist humanized antibodies) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL2/3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL4.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL5A.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR2DL5B.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR3DL1.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR3DL2.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-KIR3DL3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-NKG2A.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-NKG2C.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-NKG2E.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with IL-10.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-IL10.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anti-TSLP (thymic stromallymphopoietin).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PEGylated IL-10. In an embodiment ofthe invention, PEGylated-IL-10 is administered to the subject at a doseof up to 20 micrograms/kg (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19 or 20 micrograms/kg). For example, up to 20micrograms/kg daily, e.g., for up to four (e.g., 1, 2, 3 or 4) 28 daycycles—e.g., 20 micrograms/kg/day for four 28 day cycles.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 13-cis-retinoic acid.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with3-[5-(methylsulfonylpiperadinemethyl)-indolyl]-quinolone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 4-hydroxytamoxifen.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 5-deooxyuridine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 5′-deoxy-5-fluorouridine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 5-fluorouracil.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 6-mecaptopurine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 7-hydroxystaurosporine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with A-443654.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with abirateroneacetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with abraxane.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ABT-578.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with acolbifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ADS-100380.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ALT-110.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with altretamine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with amifostine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with aminoglutethimide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with amrubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Amsacrine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anagrelide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with anastrozole.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with angiostatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with AP-23573.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ARQ-197.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with arzoxifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with AS-252424.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with AS-605240.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with asparaginase.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with AT-9263.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with atrasentan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with axitinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with AZD1152.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Bacillus Calmette-Guerin (BCG)vaccine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with batabulin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BC-210.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with besodutox.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with bevacizumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with bicalutamide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Bio111.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BIO140.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with bleomycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BMS-214662.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BMS-247550.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BMS-275291.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BMS-310705.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with bortezimib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with buserelin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with busulfan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with calcitriol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with camptothecin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with canertinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with capecitabine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with carboplatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with carmustine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CC8490.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CEA (recombinantvaccinia-carcinoembryonic antigen vaccine).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cediranib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CG-1521.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CG-781.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with chlamydocin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with chlorambucil.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with chlorotoxin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cilengitide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cimitidine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cisplatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cladribine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with clodronate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with COL-3.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with CP-724714.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cyclophosphamide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cyproterone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cyproteroneacetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cytarabine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cytosinearabinoside.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dacarbazine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dacinostat.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dactinomycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dalotuzumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with danusertib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dasatanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with daunorubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with decatanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with deguelin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with denileukin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with deoxycoformycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with depsipeptide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with diarylpropionitrile.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with diethylstilbestrol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with diftitox.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with docetaxel.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dovitinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with doxorubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with droloxifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with edotecarin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with yttrium-90 labeled-edotreotide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with edotreotide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with EKB-569.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with EMD121974.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with endostatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with enzalutamide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with enzastaurin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with epirubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with epithilone B.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ERA-923.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Cetuximab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with erlotinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with estradiol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with estramustine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with etoposide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with everolimus.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with exemestane.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ficlatuzumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with finasteride.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with flavopiridol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with floxuridine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with fludarabine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with fludrocortisone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with fluoxymesterone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with flutamide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with FOLFOX regimen.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with fulvestrant.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with galeterone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with gefitinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with gemcitabine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with gimatecan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with glycopyranosyl lipid A.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with goserelin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with goserelin acetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with gossypol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with GSK461364.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with GSK690693.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with HMR-3339.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with hydroxyprogesteronecaproate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with hydroxyurea.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with IC87114.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with idarubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with idoxyfene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ifosfamide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with IM862.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with imatinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with imiquimod.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with IMC-1C11.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with INCB24360.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with INO1001.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with interferon.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with interleukin-2 (IL-2).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with interleukin-12 (IL-12).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ipilimumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with irinotecan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with JNJ-16241199.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ketoconazole.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with KRX-0402.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lapatinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lasofoxifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with letrozole.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with leucovorin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with leuprolide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with leuprolide acetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with levamisole.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with liposome entrapped paclitaxel.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lomustine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lonafarnib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lucanthone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY292223.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY292696.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY293646.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY293684.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY294002.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY317615.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with marimastat.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mechlorethamine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with medroxyprogesteroneacetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with megestrolacetate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with melphalan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mercaptopurine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mesna.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with methotrexate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mithramycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mitomycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mitotane.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with mitoxantrone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tozasertib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with a suspension of heat killedMycobacterium obuense.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with MLN8054.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with neovastat.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Neratinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with neuradiab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with nilotinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with nilutimide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with nolatrexe.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with NVP-BEZ235.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with oblimersen.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with octreotide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ofatumumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with oregovomab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with orteronel.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with oxaliplatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with paclitaxel.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with palbociclib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pamidronate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with panitumumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pazopanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PD0325901.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PD184352.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PEG-interferon.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pemetrexed.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pentostatin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with perifosine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with phenylalanine mustard.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PI-103.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pictilisib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PIK-75.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with pipendoxifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PKI-166.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with plicamycin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with poly-ICLC.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with porfimer.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with prednisone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with procarbazine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with progestins.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PSK protein bound polysaccharide(derived from Basidiomycete coriolus versicolor).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PX-866.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with R-763.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with raloxifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with raltitrexed.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with razoxin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ridaforolimus.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with rituximab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with romidepsin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with RTA744.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with rubitecan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with scriptaid.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Sdx102.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with seliciclib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with selumetinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with semaxanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with SF1126.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with sirolimus.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with SN36093.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with sorafenib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with spironolactone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with squalamine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with SR13668.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with streptozocin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with SU6668.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with suberoylanalide hydroxamic acid.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with sunitinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with synthetic estrogen.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with talampanel.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with talimogene laherparepvec.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tamoxifen.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with temozolomide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with temsirolimus.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with teniposide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tesmilifene.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with testosterone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tetrandrine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with TGX-221.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with thalidomide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with 6-thioguanine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with thiotepa.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ticilimumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tipifarnib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tivozanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with TKI-258.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with TLK286.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with topotecan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with toremifene citrate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with trabectedin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with trastuzumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tretinoin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with trichostatin A.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with triciribinephosphate monohydrate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with triptorelin pamoate.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with TSE-424.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tumor necrosis factor alpha (TNFα).

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with uracil mustard.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with valproic acid.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with valrubicin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vandetanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vatalanib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with VEGF trap.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vinblastine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vincristine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vindesine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vinorelbine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vitaxin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vitespan.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vorinostat.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with VX-745.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with wortmannin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Xr311.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with zanolimumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with Z-100 hot water extract of Bacillustuberculosis.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ZK186619.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ZK-304709.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ZM336372.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ZSTK474.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with casopitant.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with netupitant.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with an NK-1 receptor antagonist.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with palonosetron.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with aprepitant.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with diphenhydramine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with hydroxyzine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with metoclopramide.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with lorazepam.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with alprazolam.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with haloperidol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with droperidol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dronabinol.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dexamethasone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with methylprednisolone.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with prochlorperazine.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with granisetron.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ondansetron.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dolasetron.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with tropisetron.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with filgrastim.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PEG-filgrastim.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with erythropoietin.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with epoetin alfa.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with darbepoetin alfa.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with dabrafenib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with trametinib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with vemurafenib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with cobimetnib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LY3009120.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with DNE03.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ATI13387.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ganetespib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with encorafenib.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with MEK162.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BKM120.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with LEE011.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with BGJ398.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with INC280.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with PLX8394.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with ornatuzumab.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with natitoclax.

In an embodiment of the invention, an anti-LAG3 antibody orantigen-binding fragment thereof (e.g., humanized antibody such as anantagonist humanized antibody) of the present invention (e.g., 4A10,19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8and/or Ab9) is in association with aflibercept.

The term “in association with” indicates that the components, ananti-LAG3 antibody (e.g., humanized antibody such as antagonisthumanized antibodies) or antigen-binding fragment thereof of the presentinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) along with another agent such aspembrolizumab or nivolumab, can be formulated into a single composition,e.g., for simultaneous delivery, or formulated separately into two ormore compositions (e.g., a kit). Each component can be administered to asubject at a different time than when the other component isadministered; for example, each administration may be givennon-simultaneously (e.g., separately or sequentially) at intervals overa given period of time. Moreover, the separate components may beadministered to a subject by the same or by a different route (e.g.,wherein an anti-LAG3 antibody (e.g., humanized antibody such asantagonist humanized antibodies) or antigen-binding fragment thereof(e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8 and/or Ab9 is administered parenterally and paclitaxel isadministered orally).

Assays and Experimental and Diagnostic Uses

The present invention includes any method for forming a complex betweenan anti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention and LAG3 (e.g., human or cynomolgous monkey LAG3) comprisingcontacting the LAG3 polypeptide with the anti-LAG3 antibody or fragmentunder conditions suitable for binding and complex formation.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may be used as affinity purificationagents. In this process, the anti-LAG3 antibodies and antigen-bindingfragments thereof are immobilized on a solid phase such a sephadex,glass or agarose resin or filter paper, using methods well known in theart. The immobilized antibody or fragment is contacted with a samplecontaining the LAG3 protein (or a fragment thereof) to be purified, and,thereafter, the support is washed with a suitable solvent that willremove substantially all of the material in the sample except the LAG3protein which is bound to the immobilized antibody or fragment. Finally,the support is washed with a solvent which elutes the bound LAG3 (e.g.,protein A). Such immobilized antibodies and fragments form part of thepresent invention.

The present invention provides methods for using the anti-LAG3antibodies and antigen-binding fragments thereof of the presentinvention to determine the extent of T-cell activation that a particularsubject is having or could have in the present of the antibody orfragment. For example, embodiments of the invention include methodsincluding (i) contacting T-cells (e.g., CD4+ T-cells) from a subjectwith superantigen (e.g., any one or more of a staphylococcalsuperantigen such as SEA, SEB (Staphylococcus enterotoxin B), SEC2,SEC3, SED, SEH and/or TSST; and/or any one or more of a streptococcalsuperantigen such as SPE-A, SPE-C, SPE-H and/or SMEZ-2), e.g., at aconcentration of 500 pg/ml or more, such as about 10 ng/ml or 100 ng/ml,in the presence of the anti-LAG3 antibody or fragment (optionally, theT-cells are pre-incubated with the superantigen (e.g., SEB) and antibodyor fragment for about 48 or 72 hours) and (ii) determining the level ofproduction of cytokine (e.g., TNF-alpha, GM-CSF, IFN-gamma and/or IL-2)production of said T-cells; wherein the level of production of saidcytokine(s) indicates the level of T-cell activation in the present ofthe antibody or fragment. Subjects possessing T-cells which exhibitcytokine production (e.g., high levels of cytokine production such as ananti-LAG3-dependent increase thereof) in the presence of superantigenand antibody are considered superior candidates for receipt of theantibody or fragment as a therapy, e.g., for treating cancer orinfection. In an embodiment of the invention, such superior candidatesare selected for receipt of the antibody or fragment. In an embodimentof the invention, such superior candidates are administered an effectiveamount of the antibody or fragment. In an embodiment of the invention,the method includes the step of isolating the T-cells from the blood ofthe subject. In an embodiment of the invention, the T-cells arecontacted with anti-LAG3 antibody or antigen-binding fragment thereof ofthe present invention and pembrolizumab.

Further provided are antigens for generating secondary antibodies whichare useful, for example, for performing Western blots and otherimmunoassays discussed herein. In particular, polypeptides are disclosedwhich comprise the variable regions and/or CDR sequences of an anti-LAG3antibody or fragment disclosed herein (e.g., 4A10, 19E8, 11C9 and/or22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) and whichmay be used to generate anti-idiotypic antibodies for use inspecifically detecting the presence of the antibody, e.g., in atherapeutic context.

The present invention includes cell-based ELISA methods using theanti-LAG3 antibodies and antigen-binding fragments thereof of thepresent invention. In an embodiment of the invention, the methodincludes the steps: (i) contacting cells (e.g., cells or tissue takenfrom a tumor, e.g., which include lymphocytes suspected of expressingLAG3) immobilized to a solid surface (e.g., a microplate) to be testedfor the presence of LAG3 with an anti-LAG3 antibody or antigen-bindingfragment thereof of the present invention, (ii) optionally washing themixture to remove unbound anti-LAG3 antibody or fragment, (iii)contacting the anti-LAG3 antibody or fragment with a labeled secondaryantibody or antigen-binding fragment thereof that binds to the anti-LAG3antibody or fragment, (iv) optionally washing the complex to removeunbound antibodies or fragments and (v) detecting the presence of thelabel on the secondary antibody or fragment; wherein detection of thelabel indicates that the cells contain LAG3. For example, the presentinvention includes such cell-based ELISA methods for identifying LAG3+cells in a tumor sample.

The present invention includes ELISA assays (enzyme-linked immunosorbentassay) incorporating the use of an anti-LAG3 antibody (e.g., humanizedantibodies such as antagonist humanized antibodies) or antigen-bindingfragment thereof disclosed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9).

For example, such a method comprises the following steps:

-   (a) coat a substrate (e.g., surface of a microtiter plate well,    e.g., a plastic plate) with anti-LAG3 antibody (e.g., humanized    antibodies such as antagonist humanized antibodies) or    antigen-binding fragment thereof (e.g., 4A10, 19E8, 11C9 and/or    22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9);-   (b) apply a sample to be tested for the presence of LAG3 to the    substrate (e.g., cells taken from a tumor, e.g., which include    lymphocytes suspected of expressing LAG3);-   (c) wash the plate, so that unbound material in the sample is    removed;-   (d) apply detectably labeled antibodies (e.g., enzyme-linked    antibodies) which are also specific to the LAG3 antigen;-   (e) wash the substrate, so that the unbound, labeled antibodies are    removed;-   (f) if the labeled antibodies are enzyme linked, apply a chemical    which is converted by the enzyme into a fluorescent signal; and-   (g) detect the presence of the labeled antibody.

Detection of the label associated with the substrate indicates thepresence of the LAG3 protein. The ELISA methods can also be usedidentifying LAG3+ cells in a tumor sample.

In a further embodiment, the labeled antibody or antigen-bindingfragment thereof is labeled with peroxidase which react with ABTS (e.g.,2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)) or3,3′,5,5′-Tetramethylbenzidine to produce a color change which isdetectable. Alternatively, the labeled antibody or fragment is labeledwith a detectable radioisotope (e.g., ³H) which can be detected byscintillation counter in the presence of a scintillant.

An anti-LAG3 antibody (e.g., humanized antibodies such as antagonisthumanized antibodies) or antigen-binding fragment thereof of theinvention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may be used in a Western blot orimmune-protein blot procedure. Such a procedure forms part of thepresent invention and includes e.g.:

-   (1) providing a membrane or other solid substrate comprising a    sample to be tested for the presence of LAG3, e.g., optionally    including the step of transferring proteins from a sample to be    tested for the presence of LAG3 (e.g., from a PAGE or SDS-PAGE    electrophoretic separation of the proteins in the sample) onto a    membrane or other solid substrate using a method known in the art    (e.g., semi-dry blotting or tank blotting); and contacting the    membrane or other solid substrate to be tested for the presence of    bound LAG3 or a fragment thereof with an anti-LAG3 antibody or    antigen-binding fragment thereof of the invention.

Such a membrane may take the form, for example, of a nitrocellulose orvinyl-based (e.g., polyvinylidene fluoride (PVDF)) membrane to which theproteins to be tested for the presence of LAG3 in a non-denaturing PAGE(polyacrylamide gel electrophoresis) gel or SDS-PAGE (sodium dodecylsulfate polyacrylamide gel electrophoresis) gel have been transferred(e.g., following electrophoretic separation in the gel). Beforecontacting the membrane with the anti-LAG3 antibody or fragment, themembrane is optionally blocked, e.g., with non-fat dry milk or the likeso as to bind non-specific protein binding sites on the membrane.

-   (2) washing the membrane one or more times to remove unbound    anti-LAG3 antibody or fragment and other unbound substances; and-   (3) detecting the bound anti-LAG3 antibody or fragment.

Detection of the bound antibody or fragment indicates that the LAG3protein is present on the membrane or substrate and in the sample.Detection of the bound antibody or fragment may be by binding theantibody or fragment with a secondary antibody (an anti-immunoglobulinantibody) which is detectably labeled and, then, detecting the presenceof the secondary antibody label.

The anti-LAG3 antibodies (e.g., humanized antibodies such as antagonisthumanized antibodies) and antigen-binding fragments thereof disclosedherein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be used forimmunohistochemistry. Such a method forms part of the present inventionand comprises, e.g.,

-   (1) contacting tissue comprising TILs and tumor cells (e.g.,    melanoma tumor) to be tested for the presence of LAG3 protein with    an anti-LAG3 antibody or antigen-binding fragment thereof of the    invention; and-   (2) detecting the antibody or fragment on or in the TILs.

If the antibody or fragment itself is detectably labeled, it can bedetected directly. Alternatively, the antibody or fragment may be boundby a detectably labeled secondary antibody wherein the label is thendetected.

Certain anti-LAG3 antibodies and antigen-binding fragments thereofdisclosed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) may also be used for in vivotumor imaging. Such a method may include injection of a detectablylabeled, e.g., radiolabeled, anti-LAG3 antibody or antigen-bindingfragment thereof (as discussed herein) into the body of a patient to betested for the presence of a tumor associated with LAG3 expression(e.g., which expresses LAG3, for example, on tumor infiltratinglymphocytes (TILs)) followed by imagine, e.g., nuclear imaging, of thebody of the patient to detect the presence of the labeled antibody orfragment e.g., at loci comprising a high concentration of the antibodyor fragment which are bound to or associated with the tumor. Thedetection of the loci indicates the presence of the LAG3⁺ TILs in atumor.

Imaging techniques include SPECT imaging (single photon emissioncomputed tomography) or PET imaging (positron emission tomography).Labels include e.g., iodine-123 (¹²³I) and technetium-99m (^(99m)Tc),e.g., in conjunction with SPECT imaging or ¹¹C, ¹³N, ¹⁵O or ¹⁸F, e.g.,in conjunction with PET imaging or Indium-111 (See e.g., Gordon et al.,(2005) International Rev. Neurobiol. 67:385-440).

The present invention provide a method for determining whether a tumorin a subject is sensitive to treatment with an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention comprisingdetermining whether the LAG3 is expressed in or on the tumorinfiltrating lymphocytes (TILs) and, if said expression is identified,determining that the tumor is sensitive to said treatment. The TILs canbe determined to express LAG3 using any of the methods set forth herein,e.g., ELISA or in vivo imaging. In an embodiment of the invention, themethod comprises the step of obtaining a sample of said tumor tissuebefore making the determination of LAG3 expression is done. For example,in an embodiment of the invention, the sample is obtained surgically,e.g., by biopsy, for example, needle biopsy or partial tumorectomy. Inan embodiment of the invention, LAG3 expression is determined bycontacting the TILs with the antibody or fragment and detecting thepresence of the antibody or fragment bound to the tumor tissue orfragment.

Pharmaceutical Compositions and Administration

To prepare pharmaceutical or sterile compositions of the anti-LAG3antibodies (e.g., humanized antibodies such as antagonist humanizedantibodies) and antigen-binding fragments thereof (e.g., 4A10, 19E8,11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/orAb9), the antibody or antigen-binding fragment thereof is admixed with apharmaceutically acceptable carrier or excipient. See, e.g., Remington'sPharmaceutical Sciences and U.S. Pharmacopeia: National Formulary, MackPublishing Company, Easton, Pa. (1984). Such compositions are part ofthe present invention.

The scope of the present invention includes dessicated, e.g.,freeze-dried, compositions comprising an anti-LAG3 antibody orantigen-binding fragment thereof or a pharmaceutical composition thereofthat includes a pharmaceutically acceptable carrier but substantiallylacks water.

Formulations of therapeutic and diagnostic agents may be prepared bymixing with acceptable carriers, excipients, or stabilizers in the formof, e.g., lyophilized powders, slurries, aqueous solutions orsuspensions (see, e.g., Hardman, et al. (2001) Goodman and Gilman's ThePharmacological Basis of Therapeutics, McGraw-Hill, New York, N.Y.;Gennaro (2000) Remington: The Science and Practice of Pharmacy,Lippincott, Williams, and Wilkins, New York, N.Y.; Avis, et al. (eds.)(1993) Pharmaceutical Dosage Forms: Parenteral Medications, MarcelDekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms:Tablets, Marcel Dekker, N.Y.; Lieberman, et al. (eds.) (1990)Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, N.Y.;Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, MarcelDekker, Inc., New York, N.Y.).

The present invention includes any pharmaceutical formulation comprisingan anti-LAG3 antibody or antigen-binding fragment thereof of the presentinvention, and PO₄ ²⁻, for example, sodium phosphate or potassiumphosphate (e.g., about 10 mM), NaCl (e.g., about 7.5 mM) and sucrose(e.g., about 3%), e.g., having a pH of about 7.4 or about 7.3; or NaOAc(e.g., about 20 mM), and sucrose (about 7 or about 9%), e.g., having apH of about 5.0.

Toxicity and therapeutic efficacy of the antibody or fragmentcompositions, administered alone or in combination with anothertherapeutic agent, can be determined by standard pharmaceuticalprocedures in cell cultures or experimental animals, e.g., fordetermining the LD₅₀ (the dose lethal to 50% of the population) and theED₅₀ (the dose effective in 50% of the population). The dose ratiobetween toxic and therapeutic effects is the therapeutic index(LD₅₀/ED₅₀). In particular aspects, antibodies exhibiting hightherapeutic indices are desirable. The data obtained from these cellculture assays and animal studies can be used in formulating a range ofdosage for use in human. The dosage of such compounds lies preferablywithin a range of circulating concentrations that include the ED₅₀ withlittle or no toxicity. The dosage may vary within this range dependingupon the dosage form employed and the route of administration.

In a further embodiment, a further therapeutic agent that isadministered to a subject in association with an anti-LAG3 antibody(e.g., humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof disclosed herein (e.g., 4A10, 19E8,11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/orAb9) is administered to the subject in accordance with the Physicians'Desk Reference 2003 (Thomson Healthcare; 57th edition (Nov. 1, 2002)).

The mode of administration can vary. Routes of administration includeoral, rectal, transmucosal, intestinal, parenteral; intramuscular,subcutaneous, intradermal, intramedullary, intrathecal, directintraventricular, intravenous, intraperitoneal, intranasal, intraocular,inhalation, insufflation, topical, cutaneous, transdermal, orintra-arterial.

The present invention provided methods for administering an anti-LAG3antibody or antigen-binding fragment thereof comprising introducing theantibody or fragment into the body of a subject. For example, the methodcomprises piercing the body of the subject with a needle of a syringeand injecting the antibody or fragment into the body of the subject,e.g., into the vein, artery, tumor, muscular tissue or subcutis of thesubject.

The present invention provides a vessel (e.g., a plastic or glass vial,e.g., with a cap or a chromatography column, hollow bore needle or asyringe cylinder) comprising any of the antibodies or antigen-bindingfragments (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9), polypeptides or polynucleotides setforth herein or a pharmaceutical composition thereof comprising apharmaceutically acceptable carrier.

The present invention also provides an injection device comprising anyof the antibodies or antigen-binding fragments, polypeptides orpolynucleotides set forth herein or a pharmaceutical compositionthereof. An injection device is a device that introduces a substanceinto the body of a patient via a parenteral route, e.g., intramuscular,subcutaneous or intravenous.

For example, an injection device may be a syringe (e.g., pre-filled withthe pharmaceutical composition, such as an auto-injector) which, forexample, includes a cylinder or barrel for holding fluid to be injected(e.g., comprising the antibody or fragment or a pharmaceuticalcomposition thereof), a needle for piecing skin and/or blood vessels forinjection of the fluid; and a plunger for pushing the fluid out of thecylinder and through the needle bore. In an embodiment of the invention,an injection device that comprises an antibody or antigen-bindingfragment thereof of the present invention or a pharmaceuticalcomposition thereof is an intravenous (IV) injection device. Such adevice includes the antibody or fragment or a pharmaceutical compositionthereof in a cannula or trocar/needle which may be attached to a tubewhich may be attached to a bag or reservoir for holding fluid (e.g.,saline; or lactated ringer solution comprising NaCl, sodium lactate,KCl, CaCl₂ and optionally including glucose) introduced into the body ofthe patient through the cannula or trocar/needle. The antibody orfragment or a pharmaceutical composition thereof may, in an embodimentof the invention, be introduced into the device once the trocar andcannula are inserted into the vein of a subject and the trocar isremoved from the inserted cannula. The IV device may, for example, beinserted into a peripheral vein (e.g., in the hand or arm); the superiorvena cava or inferior vena cava, or within the right atrium of the heart(e.g., a central IV); or into a subclavian, internal jugular, or afemoral vein and, for example, advanced toward the heart until itreaches the superior vena cava or right atrium (e.g., a central venousline). In an embodiment of the invention, an injection device is anautoinjector; a jet injector or an external infusion pump. A jetinjector uses a high-pressure narrow jet of liquid which penetrate theepidermis to introduce the antibody or fragment or a pharmaceuticalcomposition thereof to a patient's body. External infusion pumps aremedical devices that deliver the antibody or fragment or apharmaceutical composition thereof into a patient's body in controlledamounts. External infusion pumps may be powered electrically ormechanically. Different pumps operate in different ways, for example, asyringe pump holds fluid in the reservoir of a syringe, and a moveablepiston controls fluid delivery, an elastomeric pump holds fluid in astretchable balloon reservoir, and pressure from the elastic walls ofthe balloon drives fluid delivery. In a peristaltic pump, a set ofrollers pinches down on a length of flexible tubing, pushing fluidforward. In a multi-channel pump, fluids can be delivered from multiplereservoirs at multiple rates.

The pharmaceutical compositions disclosed herein may also beadministered with a needleless hypodermic injection device; such as thedevices disclosed in U.S. Pat. Nos. 6,620,135; 6,096,002; 5,399,163;5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556. Suchneedleless devices comprising the pharmaceutical composition are alsopart of the present invention. The pharmaceutical compositions disclosedherein may also be administered by infusion. Examples of well-knownimplants and modules for administering the pharmaceutical compositionsinclude those disclosed in: U.S. Pat. No. 4,487,603, which discloses animplantable micro-infusion pump for dispensing medication at acontrolled rate; U.S. Pat. No. 4,447,233, which discloses a medicationinfusion pump for delivering medication at a precise infusion rate; U.S.Pat. No. 4,447,224, which discloses a variable flow implantable infusionapparatus for continuous drug delivery; U.S. Pat. No. 4,439,196, whichdiscloses an osmotic drug delivery system having multi-chambercompartments. Many other such implants, delivery systems, and modulesare well known to those skilled in the art and those comprising thepharmaceutical compositions of the present invention are within thescope of the present invention.

Alternately, one may administer the anti-LAG3 antibody (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragment (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8 and/or Ab9) in a local rather than systemic manner,for example, via injection of the antibody or fragment directly into atumor, e.g., a LAG3⁺ tumor. Furthermore, one may administer the antibodyor fragment in a targeted drug delivery system, for example, in aliposome coated with a tissue-specific antibody, targeting, for example,a tumor e.g., a LAG3⁺ tumor, e.g., characterized by immunopathology. Theliposomes will be targeted to and taken up selectively by the afflictedtissue. Such methods and liposomes are part of the present invention.

“Treat” or “treating” means to administer anti-LAG3 antibodies orantigen-binding fragments thereof of the present invention, to a subjecthaving one or more symptoms of a disease for which the anti-LAG3antibodies and antigen-binding fragments are effective, e.g., in thetreatment of a subject having cancer or an infectious disease, or beingsuspected of having cancer or infectious disease, for which the agenthas therapeutic activity. Typically, the antibody or fragment isadministered in an “effective amount” or “effective dose” which willalleviate one or more symptoms (e.g., of cancer or infectious disease)in the treated subject or population, whether by inducing the regressionor elimination of such symptoms or by inhibiting the progression of suchsymptom(s), e.g., cancer symptoms such as tumor growth or metastasis, byany clinically measurable degree. The effective amount of the antibodyor fragment may vary according to factors such as the disease stage,age, and weight of the patient, and the ability of the drug to elicit adesired response in the subject.

Antibodies (e.g., humanized antibodies such as antagonist humanizedantibodies) or antigen-binding fragments thereof disclosed herein (e.g.,4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7,Ab8 and/or Ab9) may be provided by continuous infusion, or by dosesadministered, e.g., daily, 1-7 times per week, weekly, bi-weekly,monthly, bimonthly, quarterly, semiannually, annually etc. Doses may beprovided, e.g., intravenously, subcutaneously, topically, orally,nasally, rectally, intramuscular, intracerebrally, intraspinally, or byinhalation. An effective dose of an anti-LAG3 antibody orantigen-binding fragment thereof of the present invention, is from about0.1 mg/kg (body weight) to about 100 mg/kg (body weight), e.g., fortreatment or prevention of cancer or infectious diseases. In anembodiment of the invention, an effective dose for treatment of amedical condition, wherein overexpression of LAG3 occurs, is the dose atwhich complete saturation of the LAG3 antigen occurs in the body of thesubject, e.g., on T-cells within tumors of the subject or wherein thereis about 10% saturation or more than about 10%.

Determination of the appropriate dose is made by the clinician, e.g.,using parameters or factors known or suspected in the art to affecttreatment. Generally, in determining the dose, the dose begins with anamount somewhat less than the optimum dose and it is increased by smallincrements thereafter until the desired or optimum effect is achievedrelative to any negative side effects. Important diagnostic measuresinclude those of symptoms of, e.g., the inflammation or level ofinflammatory cytokines produced. In general, it is desirable that abiologic that will be used is derived from the same species as theanimal targeted for treatment, thereby minimizing any immune response tothe reagent. In the case of human subjects, for example, chimeric,humanized and fully human antibodies are may be desirable. Guidance inselecting appropriate doses of anti-LAG3 antibodies or fragments isavailable (see, e.g., Wawrzynczak (1996) Antibody Therapy, BiosScientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) MonoclonalAntibodies, Cytokines and Arthritis, Marcel Dekker, New York, N.Y.; Bach(ed.) (1993) Monoclonal Antibodies and Peptide Therapy in AutoimmuneDiseases, Marcel Dekker, New York, N.Y.; Baert et al. (2003) New Engl.J. Med. 348:601-608; Milgrom et al. (1999) New Engl. J. Med.341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792;Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al.(2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J.Med. 343:1594-1602).

Whether a disease symptom has been alleviated can be assessed by anyclinical measurement typically used by physicians or other skilledhealthcare providers to assess the severity or progression status ofthat symptom. While an embodiment of the present invention (e.g., atreatment method or article of manufacture) may not be effective inalleviating the target disease symptom(s) in every subject, it shouldalleviate the target disease symptom(s) in a statistically significantnumber of subjects as determined by any statistical test known in theart such as the Student's t-test, the chi²-test, the U-test according toMann and Whitney, the Kruskal-Wallis test (H-test),Jonckheere-Terpstra-test and the Wilcoxon-test.

Kits

Further provided are kits comprising one or more components thatinclude, but are not limited to, an anti-LAG3 antibody (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragment, as discussed herein (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g.,Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) in association withone or more additional components including, but not limited to, afurther therapeutic agent, as discussed herein. The antibody or fragmentand/or the therapeutic agent can be formulated as a pure composition orin combination with a pharmaceutically acceptable carrier, in apharmaceutical composition.

In one embodiment, the kit includes an anti-LAG3 antibody (e.g.,humanized antibody such as antagonist humanized antibodies) orantigen-binding fragment thereof of the invention (e.g., 4A10, 19E8,11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/orAb9) or a pharmaceutical composition thereof in one container (e.g., ina sterile glass or plastic vial) and a further therapeutic agent inanother container (e.g., in a sterile glass or plastic vial).

In another embodiment, the kit comprises a combination of the invention,including an anti-LAG3 antibody (e.g., humanized antibody such asantagonist humanized antibodies) or antigen-binding fragment thereof ofthe invention (e.g., 4A10, 19E8, 11C9 and/or 22D2; e.g., Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) or pharmaceutical compositionthereof in combination with one or more therapeutic agents formulatedtogether, optionally, in a pharmaceutical composition, in a single,common container.

If the kit includes a pharmaceutical composition for parenteraladministration to a subject, the kit can include a device for performingsuch administration. For example, the kit can include one or morehypodermic needles or other injection devices as discussed above. Thus,the present invention includes a kit comprising an injection device andthe anti-LAG3 antibody or antigen-binding fragment thereof of thepresent invention, e.g., wherein the injection device includes theantibody or fragment or wherein the antibody or fragment is in aseparate vessel.

The kit can include a package insert including information concerningthe pharmaceutical compositions and dosage forms in the kit. Generally,such information aids patients and physicians in using the enclosedpharmaceutical compositions and dosage forms effectively and safely. Forexample, the following information regarding a combination of theinvention may be supplied in the insert: pharmacokinetics,pharmacodynamics, clinical studies, efficacy parameters, indications andusage, contraindications, warnings, precautions, adverse reactions,overdosage, proper dosage and administration, how supplied, properstorage conditions, references, manufacturer/distributor information andpatent information.

Detection Kits and Therapeutic Kits

As a matter of convenience, an anti-LAG3 antibody (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragment thereof of the invention (e.g., 4A10, 19E8, 11C9 and/or 22D2;e.g., Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 and/or Ab9) can be providedin a kit, i.e., a packaged combination of reagents in predeterminedamounts with instructions for performing the diagnostic or detectionassay. Where the antibody or fragment is labeled with an enzyme, the kitwill include substrates and cofactors required by the enzyme (e.g., asubstrate precursor which provides the detectable chromophore orfluorophore). In addition, other additives may be included such asstabilizers, buffers (e.g., a block buffer or lysis buffer) and thelike. The relative amounts of the various reagents may be varied widelyto provide for concentrations in solution of the reagents whichsubstantially optimize the sensitivity of the assay. Particularly, thereagents may be provided as dry powders, usually lyophilized, includingexcipients which on dissolution will provide a reagent solution havingthe appropriate concentration.

Also provided are diagnostic or detection reagents and kits comprisingone or more such reagents for use in a variety of detection assays,including for example, immunoassays such as ELISA (sandwich-type orcompetitive format). The kit's components may be pre-attached to a solidsupport, or may be applied to the surface of a solid support when thekit is used. In some embodiments of the invention, the signal generatingmeans may come pre-associated with an antibody or fragment of theinvention or may require combination with one or more components, e.g.,buffers, antibody-enzyme conjugates, enzyme substrates, or the like,prior to use. Kits may also include additional reagents, e.g., blockingreagents for reducing nonspecific binding to the solid phase surface,washing reagents, enzyme substrates, and the like. The solid phasesurface may be in the form of a tube, a bead, a microtiter plate, amicrosphere, or other materials suitable for immobilizing proteins,peptides, or polypeptides. In particular aspects, an enzyme thatcatalyzes the formation of a chemilluminescent or chromogenic product orthe reduction of a chemilluminescent or chromogenic substrate is acomponent of the signal generating means. Such enzymes are well known inthe art. Kits may comprise any of the capture agents and detectionreagents described herein. Optionally the kit may also compriseinstructions for carrying out the methods of the invention.

Also provided is a kit comprising an anti-LAG3 antibody (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragment thereof packaged in a container, such as a vial or bottle, andfurther comprising a label attached to or packaged with the container,the label describing the contents of the container and providingindications and/or instructions regarding use of the contents of thecontainer to treat one or more disease states as described herein.

In one aspect, the kit is for treating cancer and comprises an anti-LAG3antibody (e.g., humanized antibody such as antagonist humanizedantibodies) or antigen-binding fragment thereof and a furthertherapeutic agent or a vaccine. The kit may optionally further include asyringe for parenteral, e.g., intravenous, administration. In anotheraspect, the kit comprises an anti-LAG3 antibody (e.g., humanizedantibody such as antagonist humanized antibodies) or antigen-bindingfragment thereof and a label attached to or packaged with the containerdescribing use of the antibody or fragment with the vaccine or furthertherapeutic agent. In yet another aspect, the kit comprises the vaccineor further therapeutic agent and a label attached to or packaged withthe container describing use of the vaccine or further therapeutic agentwith the anti-LAG3 antibody or fragment. In certain embodiments, ananti-LAG3 antibody and vaccine or further therapeutic agent are inseparate vials or are combined together in the same pharmaceuticalcomposition.

As discussed above in the combination therapy section, concurrentadministration of two therapeutic agents does not require that theagents be administered at the same time or by the same route, as long asthere is an overlap in the time period during which the agents areexerting their therapeutic effect. Simultaneous or sequentialadministration is contemplated, as is administration on different daysor weeks.

The therapeutic and detection kits disclosed herein may also be preparedthat comprise at least one of the antibody, peptide, antigen-bindingfragment, or polynucleotide disclosed herein and instructions for usingthe composition as a detection reagent or therapeutic agent. Containersfor use in such kits may typically comprise at least one vial, testtube, flask, bottle, syringe or other suitable container, into which oneor more of the detection and/or therapeutic composition(s) may beplaced, and preferably suitably aliquoted. Where a second therapeuticagent is also provided, the kit may also contain a second distinctcontainer into which this second detection and/or therapeuticcomposition may be placed. Alternatively, a plurality of compounds maybe prepared in a single pharmaceutical composition, and may be packagedin a single container means, such as a vial, flask, syringe, bottle, orother suitable single container. The kits disclosed herein will alsotypically include a means for containing the vial(s) in closeconfinement for commercial sale, such as, e.g., injection or blow-moldedplastic containers into which the desired vial(s) are retained. Where aradiolabel, chromogenic, fluorigenic, or other type of detectable labelor detecting means is included within the kit, the labeling agent may beprovided either in the same container as the detection or therapeuticcomposition itself, or may alternatively be placed in a second distinctcontainer means into which this second composition may be placed andsuitably aliquoted. Alternatively, the detection reagent and the labelmay be prepared in a single container means, and in most cases, the kitwill also typically include a means for containing the vial(s) in closeconfinement for commercial sale and/or convenient packaging anddelivery.

A device or apparatus for carrying out the detection or monitoringmethods described herein is also provided. Such an apparatus may includea chamber or tube into which sample can be input, a fluid handlingsystem optionally including valves or pumps to direct flow of the samplethrough the device, optionally filters to separate plasma or serum fromblood, mixing chambers for the addition of capture agents or detectionreagents, and optionally a detection device for detecting the amount ofdetectable label bound to the capture agent immunocomplex. The flow ofsample may be passive (e.g., by capillary, hydrostatic, or other forcesthat do not require further manipulation of the device once sample isapplied) or active (e.g., by application of force generated viamechanical pumps, electroosmotic pumps, centrifugal force, or increasedair pressure), or by a combination of active and passive forces.

In further embodiments, also provided is a processor, a computerreadable memory, and a routine stored on the computer readable memoryand adapted to be executed on the processor to perform any of themethods described herein. Examples of suitable computing systems,environments, and/or configurations include personal computers, servercomputers, hand-held or laptop devices, multiprocessor systems,microprocessor-based systems, set top boxes, programmable consumerelectronics, network PCs, minicomputers, mainframe computers,distributed computing environments that include any of the above systemsor devices, or any other systems known in the art.

General Methods

Standard methods in molecular biology are described Sambrook, Fritschand Maniatis (1982 & 1989 2^(nd) Edition, 2001 3^(rd) Edition) MolecularCloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y.; Sambrook and Russell (2001) Molecular Cloning,3^(rd) ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y.; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego,Calif.). Standard methods also appear in Ausbel, et al. (2001) CurrentProtocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. NewYork, N.Y., which describes cloning in bacterial cells and DNAmutagenesis (Vol. 1), cloning in mammalian cells and yeast (Vol. 2),glycoconjugates and protein expression (Vol. 3), and bioinformatics(Vol. 4).

Methods for protein purification including immunoprecipitation,chromatography, electrophoresis, centrifugation, and crystallization aredescribed (Coligan, et al. (2000) Current Protocols in Protein Science,Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis,chemical modification, post-translational modification, production offusion proteins, glycosylation of proteins are described (see, e.g.,Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2,John Wiley and Sons, Inc., New York; Ausubel, et al. (2001) CurrentProtocols in Molecular Biology, Vol. 3, John Wiley and Sons, Inc., NY,N.Y., pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for LifeScience Research, St. Louis, Mo.; pp. 45-89; Amersham Pharmacia Biotech(2001) BioDirectory, Piscataway, N.J., pp. 384-391). Production,purification, and fragmentation of polyclonal and monoclonal antibodiesare described (Coligan, et al. (2001) Current Protcols in Immunology,Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999)Using Antibodies, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y.; Harlow and Lane, supra). Standard techniques forcharacterizing ligand/receptor interactions are available (see, e.g.,Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, JohnWiley, Inc., New York).

Monoclonal, polyclonal, and humanized antibodies can be prepared (see,e.g., Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ.Press, New York, N.Y.; Kontermann and Dubel (eds.) (2001) AntibodyEngineering, Springer-Verlag, New York; Harlow and Lane (1988)Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press,Cold Spring Harbor, N.Y., pp. 139-243; Carpenter, et al. (2000) J.Immunol. 165:6205; He, et al. (1998) J. Immunol. 160:1029; Tang et al.(1999) J. Biol. Chem. 274:27371-27378; Baca et al. (1997) J. Biol. Chem.272:10678-10684; Chothia et al. (1989) Nature 342:877-883; Foote andWinter (1992) J. Mol. Biol. 224:487-499; U.S. Pat. No. 6,329,511).

An alternative to humanization is to use human antibody librariesdisplayed on phage or human antibody libraries in transgenic mice(Vaughan et al. (1996) Nature Biotechnol. 14:309-314; Barbas (1995)Nature Medicine 1:837-839; Mendez et al. (1997) Nature Genetics15:146-156; Hoogenboom and Chames (2000) Immunol. Today 21:371-377;Barbas et al. (2001) Phage Display: A Laboratory Manual, Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y.; Kay et al. (1996)Phage Display of Peptides and Proteins: A Laboratory Manual, AcademicPress, San Diego, Calif.; de Bruin et al. (1999) Nature Biotechnol.17:397-399).

Single chain antibodies and diabodies are described (see, e.g., Maleckiet al. (2002) Proc. Natl. Acad. Sci. USA 99:213-218; Conrath et al.(2001) J. Biol. Chem. 276:7346-7350; Desmyter et al. (2001) J. Biol.Chem. 276:26285-26290; Hudson and Kortt (1999) J. Immunol. Methods231:177-189; and U.S. Pat. No. 4,946,778). Bifunctional antibodies areprovided (see, e.g., Mack, et al. (1995) Proc. Natl. Acad. Sci. USA92:7021-7025; Carter (2001) J. Immunol. Methods 248:7-15; Volkel, et al.(2001) Protein Engineering 14:815-823; Segal, et al. (2001) J. Immunol.Methods 248:1-6; Brennan, et al. (1985) Science 229:81-83; Raso, et al.(1997) J. Biol. Chem. 272:27623; Morrison (1985) Science 229:1202-1207;Traunecker, et al. (1991) EMBO J. 10:3655-3659; and U.S. Pat. Nos.5,932,448, 5,532,210, and 6,129,914).

Bispecific antibodies are also provided (see, e.g., Azzoni et al. (1998)J. Immunol. 161:3493; Kita et al. (1999) J. Immunol. 162:6901; Merchantet al. (2000) J. Biol. Chem. 74:9115; Pandey et al. (2000) J. Biol.Chem. 275:38633; Zheng et al. (2001) J. Biol Chem. 276:12999; Propst etal. (2000) J. Immunol. 165:2214; Long (1999) Ann. Rev. Immunol. 17:875).

Purification of antigen is not necessary for the generation ofantibodies. Animals can be immunized with cells bearing the antigen ofinterest. Splenocytes can then be isolated from the immunized animals,and the splenocytes can fused with a myeloma cell line to produce ahybridoma (see, e.g., Meyaard et al. (1997) Immunity 7:283-290; Wrightet al. (2000) Immunity 13:233-242; Preston et al., supra; Kaithamana etal. (1999) J. Immunol. 163:5157-5164).

Antibodies can be conjugated, e.g., to small drug molecules, enzymes,liposomes, polyethylene glycol (PEG). Antibodies are useful fortherapeutic, diagnostic, kit or other purposes, and include antibodiescoupled, e.g., to dyes, radioisotopes, enzymes, or metals, e.g.,colloidal gold (see, e.g., Le Doussal et al. (1991) J. Immunol.146:169-175; Gibellini et al. (1998) J. Immunol. 160:3891-3898; Hsingand Bishop (1999) J. Immunol. 162:2804-2811; Everts et al. (2002) J.Immunol. 168:883-889).

Methods for flow cytometry, including fluorescence activated cellsorting (FACS), are available (see, e.g., Owens, et al. (1994) FlowCytometry Principles for Clinical Laboratory Practice, John Wiley andSons, Hoboken, N.J.; Givan (2001) Flow Cytometry, 2^(nd) ed.;Wiley-Liss, Hoboken, N.J.; Shapiro (2003) Practical Flow Cytometry, JohnWiley and Sons, Hoboken, N.J.). Fluorescent reagents suitable formodifying nucleic acids, including nucleic acid primers and probes,polypeptides, and antibodies, for use, e.g., as diagnostic reagents, areavailable (Molecular Probes (2003) Catalogue, Molecular Probes, Inc.,Eugene, Oreg.; Sigma-Aldrich (2003) Catalogue, St. Louis, Mo.).

Standard methods of histology of the immune system are described (see,e.g., Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology andPathology, Springer Verlag, New York, N.Y.; Hiatt, et al. (2000) ColorAtlas of Histology, Lippincott, Williams, and Wilkins, Phila, Pa.;Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, NewYork, N.Y.).

Software packages and databases for determining, e.g., antigenicfragments, leader sequences, protein folding, functional domains,glycosylation sites, and sequence alignments, are available (see, e.g.,GenBank, Vector NTI® Suite (Informax, Inc, Bethesda, Md.); GCG WisconsinPackage (Accelrys, Inc., San Diego, Calif.); DeCypher® (TimeLogic Corp.,Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16: 741-742;Menne, et al. (2000) Bioinformatics Applications Note 16:741-742; Wren,et al. (2002) Comput. Methods Programs Biomed. 68:177-181; von Heijne(1983) Eur. J. Biochem. 133:17-21; von Heijne (1986) Nucleic Acids Res.14:4683-4690).

EXAMPLES

These examples are intended to exemplify the present invention are not alimitation thereof. Compositions and methods set forth in the Examplesform part of the present invention.

Example 1 Generation of Antibodies Against Human LAG3

To generate antibodies to human LAG3, Balb/C mice were immunized with 5ug of human LAG3-hFc tagged recombinant protein using RIBI adjuvant andfootpad injection on a biweekly schedule. Immunized mice were bled andserum titers determined for binding to human LAG3 transfected CHOK1cells using a cell-based ELISA (described below). Mice with the highesttiters were given a final boost with recombinant protein and drainingpopliteal lymph nodes isolated four days later. Hybridomas weregenerated by electrofusion of isolated lymphocytes with the myelomafusion partner P3X63-AG8.653 using the Cytopulse Hybrimmuneelectrofusion system. Fused cells were plated in 96-well plates inDMEM/F12, 15% BCS, HAT, IL-6, OPI supplement, and gentamycin.

Hybridoma supernatants were assayed for binding to human LAG3 expressingCHOK1 cells and cross-reactivity to cynomolgus LAG3 expressing CHO cellsusing a cell-based ELISA format. Human LAG3 and cynomolgus LAG3expressing CHO-K1 cells were plated in 96-well tissue-culture plates in50u1 of DMEM/F12, 10% BCS and gentamycin (CHO-K1 media). Cells wereplated at either 2×10⁴ cells/well two days prior to the assay or 4×10⁴cells/well one day prior to the assay. Media was removed from the wellsprior to the assay and 50 ul of hybridoma supernatant added. Hybridomasupernatants were incubated for 30-60 minutes at 37° C. and washed 3times with PBS/0.05% Tween 20 using a cell ELISA washing protocol on theBiotek EL405x Select CW plate washer. Fifty microliters of the detectionantibody, HRP-conjugated goat anti-mouse IgG (Southern Biotech cat#1031-05), was added at a 1:2000 dilution in CHO-K1 media and incubatedat 37° C. for 30-60 minutes. Assay plates were washed as above anddeveloped with TMB and stopped with TMB stop solution (KPL cat#50-85-06). The absorbance at 450 nm-620 nm was determined.

Positive hybridomas were subcloned by limiting dilution or subcloned byplating hybridomas in semi-solid media and clones picked on the ClonePix(Genetix). Final subclones were grown in small-scale cultures inserum-free hybridoma production medium and purified with protein G spincolumns to generate purified antibody for further characterization.Purified antibodies from clones LB145.22D2.E1.1D1 (22D2),LB148.19E8.G1.1A1 (19E8), LB148.4A10.1H1 (4A10), and LB148.11C9.1C1(11C9) were further characterized and V_(H) and V_(L) sequencingperformed.

Example 2 Binding of Mouse Anti-LAG3 Antibodies to Human LAG3 andCynomolgous Monkey LAG3 Expressed on the Surface of Chinese HamsterOvary Cells

Mouse anti-human LAG3 antibodies were tested for binding to human LAG3and cynomolgus LAG3 expressing CHO-K1 cells using a cell-based ELISA.CHO-K1 cells were plated as described above and media removed prior toadding the test samples. Purified antibody from clones LB145.22D2.E1.1D1(22D2), LB148.19E8.G1.1A1 (19E8), LB148.4A10.1H1 (4A10), andLB148.11C9.1C1 (11C9) were serially-diluted in DMEM/F12, 10% BCS (CHOK1media) and added to the CHO-K1 plates. The samples were incubated at 37°C. for 30-60 minutes and plates were washed three times with PBS/0.05%Tween-20 using the cell wash program on the Biotek washer as describedabove. Binding was detected using an HRP-conjugated goat anti-mouse IgG(Southern Biotech cat #1031-05) secondary antibody added at a 1:2000dilution in CHO-K1 media and incubated at 37° C. for 30-60 minutes.Assay plates were washed as above and developed with TMB and stoppedwith TMB stop solution (KPL cat #50-85-06). The absorbance at 450 nm-620nm was determined. Representative binding curves for binding to humanand cynomolgus LAG3 transfected CHO-K1 cells are in FIG. 1.

Example 3 Affinity Determination for Binding of Mouse Anti-LAG3Antibodies to Human LAG3 Recombinant Protein

The kinetic binding activity of mouse anti-human LAG3 clonesLB148.19E8.G1.1A1, LB148.4A10.1H1, LB148.11C9.1C1 and LB145.22D2.E1.1D1using human LAG3-His tagged recombinant protein was measured by surfaceplasmon resonance using a Biacore T200 system (Biacore, GE Healthcare,Piscataway, N.J.). Approximately 4000 RU of Goat Anti-Mouse IgG Fcgamma, Fragment Specific (Jackson ImmunoResearch Catalog #115-006-071,Lot 81313) was immobilized via amine coupling chemistry onto a Series SCM4 sensor chip, catalog number BR-1005-34. Mouse anti-human LAG3 cloneslisted above were injected over the immobilized goat anti-mouse surfacesat 1 ug/mL for a capture level of 40 RU. HBS-EP+ buffer (BR-1006-69) wasused as the running buffer with a flow rate of 30 μL/min.

Varying concentrations of human LAG3-His protein ranging from 0.15 nM to18.8 nM, at a flow rate of 40 μL/min were injected over the antibodysurfaces. Following each injection cycle, the Series S CM4 chip surfacewas regenerated using one six second injection of 10 mM Glycine pH 1.5solution followed by an injection of 12.5 mM NaOH solution at a flowrate of 60 μL/min.

Background subtraction binding sensorgrams were used for analyzing therate constant of association (k_(a)) and dissociation (k_(d)), and theequilibrium dissociation constant K_(D). The resulting data sets werefitted with a 1:1 Langmuir Binding Model using the Biacore T200evaluation software (version 2.0). Table 4 summarizes the affinities forthe mouse anti-human LAG3 antibodies to recombinant human LAG3.

TABLE 4 Measurement of Affinity for Mouse anti-Human LAG3 Antibodies toRecombinant Human LAG3. ka1 kd1 K_(D) Clone ID (1/Ms) (1/s) (M)LB145.22D2.E1.1D1 1.39E+07 3.84E−05 2.77E−12 LB148.19E8.G1.1A1 7.43E+061.09E−04 1.47E−11 LB148.11C9.1C1 1.31E+06 1.92E−03 1.47E−09LB148.4A10.1H1 1.25E+06 1.13E−04 9.03E−11

Moreover, Ab6 binding to human CD4, which is structurally related toLAG3, both having four extracellular Ig-like domains, was not detectedby BiaCore of by FACS when CD4 was expressed on transfected L-cells.

Example 4 Effect of Anti-LAG3 Antibodies on Murine T-Cell Hybridoma 3A9Cells Expressing Human LAG3

The ability of anti-LAG3 antibodies to enhance antigen-specific T cellactivation was tested in a modified version of a previously described Tcell activation assay (Workman et al., (2002) Eur. J. Immunol.32:2255-2263).

A HEL peptide₄₈₋₆₃-specific mouse T cell hybridoma (3A9) was stimulatedwith a haplotype-matched, MHCII⁺, HEL peptide₄₈₋₆₃-pulsed B cell line(LK35.2) and IL-2 release was assessed as a readout for antigen-specificT cell activation. The 3A9 T cell response to HEL peptide₄₈₋₆₃-pulsedLK35.2 cells was dose-dependent.

3A9 T cell lines stably overexpressing mouse or human LAG3 weregenerated by retroviral transduction. We demonstrated that mouse MHC2 onLK35.2 cells can engage both human and mouse LAG3, resulting in a strongreduction of the antigen-specific IL-2 production, at suboptimal T cellactivation concentrations. The maximal effect of LAG3 activity wasobserved when titrating HEL peptide₄₈₋₆₃ at a concentration of 31.2 nM.The inhibitory effect of LAG3 overexpression was not seen when usingLK35.2 B cells pulsed with higher peptide doses (corresponding to >100nM). Treating with 10 ug/ml of a commercially available mouse LAG3antibody, C9B7W, we were able to rescue IL-2 to levels of the vector 3A9cells.

To assess the effect of anti-LAG3 antibodies in this assay, mouse orhuman LAG3-overexpressing 3A9 T cells (100,000 per well) were pretreatedwith anti-LAG3 antibodies (serially diluted in 3-fold dilutions from 10ug/ml) for 30 minutes at 37° C., and stimulated with LK35.2 cells(33,333 per well) pulsed for 30 minutes prior to co-culture with 31.2 nMHEL peptide₄₈₋₆₃. After stimulation for 24 h at 37° C. and 5.0% CO₂,IL-2 secretion was assessed in culture supernatants by mesoscale.Inhibition of LAG3 with an antagonist antibody restored T-cell functionresulting in the rescue of IL-2 production in a dose-dependent manner.IL-2 production was not rescued when 3A9 cell were pre-treated withisotype control antibodies. The ability of LAG3 overexpression tosuppress IL-2 secretion coupled with IL-2 rescue after treatment withanti-LAG3 antibody validates this assay as a robust screening tool.Table 5 lists the EC50s for IL-2 rescue using the hLAG3-3A9 system forthe mouse anti-human LAG3 antibodies.

TABLE 5 Mouse anti-human LAG3 antibodies stimulate IL-2 production inthe hLAG3-3A9 T cell system Clone ID EC50 (nM) EC50 (nM)LB145.22D2.E1.1D1 1.06 1.65 LB148.19E8.G1.1A1 1.74 1.83 LB148.11C9.1C13.56 4.06 LB148.4A10.1H1 2.83 2.96

Example 5 Blocking of LAG3/MHC Class II Binding on Daudi Cells

Mouse anti-human LAG3 clones were tested for their ability to blockhLAG3 interaction with human MHC Class II. Daudi cells (ATCC #CCL-213)were used as a cell line positive for human MHC class II expression.Daudi cells were blocked with 10 ug/ml of goat IgG in HBSS and 2% BCS onice for 30 minutes and 0.5×10⁶ cells/sample were aliquoted into a96-well V-bottom plate and blocking buffer removed. ClonesLB145.22D2.E1.1D1, LB148.19E8.G1.1A1, LB148.4A10.1H1, and LB148.11C9.1C1were serially diluted starting at 20 ug/ml in HBSS/2% BCS andpre-incubated with 2 ug/ml of human LAG3-huFc in 96-well polypropyleneU-bottom plates in a final volume of 100 ul and incubated on ice for 30minutes. Following pre-incubation, the human LAG3-Fc/hybridomasupernatant mixtures were added to the blocked Daudi cells and incubatedfor 45 minutes on ice. Cells were pelleted by centrifugation at 1200 rpmand washed twice with HBSS/2% BCS. Human LAG3-Fc binding to Daudi cellswas detected using F(ab)′₂ goat anti-human IgG-PE conjugate (SouthernBiotech Cat #) at 1:200 dilution in 100 ul staining volume and incubatedon ice for 20 minutes. Cells were washed twice as described above,resuspended in HBSS/2% BCS and read on the FACSCalibur. Table 6summarizes the IC50s for MHC class II blockade for the mouse anti-humanLAG3 clones.

TABLE 6 Mouse anti-human LAG3 antibodies block the interaction of humanMHC Class II with human LAG3-Fc recombinant protein. Clone ID IC50 (nM)LB145.22D2.E1.1D1 2.10 LB148.19E8.G1.1A1 2.80 LB148.11C9.1C1 2.00LB148.4A10.1H1 1.90

Example 6 Evaluation of Four Anti-Human LAG3 Parental Antibodies forTissue Cross-Reactivity in Focused Set of Normal Human Tissues byImmunohistochemistry

Frozen sections of a subset of normal human tissues (brain, heart,kidney, liver, lung, pancreas, pituitary) were stained using fouranti-human LAG3 antibody clones (LB148.4A10.1H1 (4A10), LB148.11C9.1C1(11C9), LB148.19E8.G1.1A1 (19E8), LB145.22D2. E1.1D1 (22D2)) asimmunohistochemical reagents, in order to screen for potentialunexpected tissue reactivity. Human tonsil was used as a positivestaining control. Mouse IgG2a, IgG1, and IgG2b, hereafter “isotypecontrol” antibodies, were run concurrent with the respective anti-humanLAG3 clones on all tissues to serve as comparators for evaluation ofnon-specific labeling.

Immunohistochemical cross-reactivity testing of antibodies 4A10, 11C9,and 19E8 and 22D2 was performed in separate runs.

All slides were reviewed by a pathologist and immunohistochemical signalwas scored on a 0-3 scale (0=negative, +1=low, +2=moderate, +3=high).Staining intensities and patterns were compared between test article andisotype control reagents. Test article staining was considered to besignificant when the intensity substantially exceeded that of theisotype control or had a distinct, reproducible difference indistributional pattern within the tissue.

TABLE 7 Lag3 clone 4A10 (5 ug/mL) Tissue Identification Test IgG2aSlide# Antibody (Human) antibody isotype Lag3 clone 4A10 5 ug/mL D6414B1 brain +1 +1 Lag3 clone 4A10 5 ug/mL D1106 B1 heart +1 +1 Lag3 clone4A10 5 ug/mL D7462 B1 kidney +1 +1 Lag3 clone 4A10 5 ug/mL D7122 B1liver +2 +2 Lag3 clone 4A10 5 ug/mL D7092 B1 lung +1 +1 Lag3 clone 4A105 ug/mL D5425 B1 pancreas +2, exocrine 0 cells Lag3 clone 4A10 5 ug/mLD5745 B1 pituitary +1 +1 Positive Control Lag3 clone 4A10 5 ug/mL D7530B2 tonsil +2 +1 (Appropriate labeling)Positive labeling of pancreatic exocrine cells (approximately 20% oftotal population present) is only seen in the test antibody treatedsample but is limited to the cytoplasm.

TABLE 8 Lag3 clone 11C9 (10 ug/mL) Tissue Identification Test IgG1Slide# Antibody (Human) antibody isotype Lag3 clone 11C9 10 ug/mL D6414B1 brain +1 +1 Lag3 clone 11C9 10 ug/mL D1106 B1 heart 0 0 Lag3 clone11C9 10 ug/mL D7462 B1 kidney 0 0 Lag3 clone 11C9 10 ug/mL D7122 B1liver +1 +2 Lag3 clone 11C9 10 ug/mL D7092 B1 lung 0 0 Lag3 clone 11C910 ug/mL D5425 B1 pancreas 0 0 Lag3 clone 11C9 10 ug/mL D5745 B1pituitary 0 0 Positive Control Lag3 clone 11C9 10 ug/mL D7530 B2 tonsil+2 0 (Appropriate labeling)All positive signal cited in tables (except in positive controls) isinterpreted as artifactually associated with the procedure performed andis identical to, or stronger in the isotype control.

TABLE 9 Lag3 clone 19E8 (0.5 ug/mL) Tissue Identification IgG2b Slide#Antibody (Human) Test antibody isotype Lag3 clone 19E8 0.5 ug/mL D6414B1 brain +1 (vascular smooth muscle) 0 Lag3 clone 19E8 0.5 ug/mL D1106B1 heart +1 (vascular smooth muscle) 0 Lag3 clone 19E8 0.5 ug/mL D7462B1 kidney +1 (vascular smooth muscle) +1 Lag3 clone 19E8 0.5 ug/mL D7122B1 liver +1 (vascular smooth muscle) +1 Lag3 clone 19E8 0.5 ug/mL D7092B1 lung 0 +1 Lag3 clone 19E8 0.5 ug/mL D5425 B1 pancreas +1 (vascularsmooth muscle) 0 Lag3 clone 19E8 0.5 ug/mL D5745 B1 pituitary +1(vascular smooth muscle) 0 Positive Control Lag3 clone 19E8 0.5 ug/mLD7530 B2 tonsil +3 (Appropriate labeling) +1Nonspecific labeling of vascular smooth muscle is only observed in thetest antibody treated samples but is limited to sarcoplasm/cytoplasm.

TABLE 10 Lag3 clone 22D2 (10 ug/mL) Tissue Identification Test IgG2aAntibody (Human) antibody isotype Lag3 clone 22D2 10 ug/mL D6414 B1brain +2 +2 Lag3 clone 22D2 10 ug/mL D1106 B1 heart +1 +1 Lag3 clone22D2 10 ug/mL D7462 B1 kidney +2 +2 Lag3 clone 22D2 10 ug/mL D7122 B1liver +1 +2 Lag3 clone 22D2 10 ug/mL D7092 B1 lung +2 +2 Lag3 clone 22D210 ug/mL D5425 B1 pancreas +1 +1 Lag3 clone 22D2 10 ug/mL D5745 B1pituitary +2 +2 Positive Control Lag3 clone 22D2 10 ug/mL D7530 B2tonsil +3 +2 (Appropriate labeling)All positive signal cited in tables (except in positive controls) isinterpreted as artifactually associated with the procedure performed andis identical to, or stronger in the isotype control.

In conclusion, clone 19E8 exhibited relatively prominent sarcoplasmiclabeling of vascular smooth muscle in all tissues examined except lung.Clone 4A10 exhibited cytoplasmic immunohistochemical reactivity inapproximately 20% of pancreatic exocrine glandular cells. Clones 11C9and 22D2 demonstrated no labeling beyond that observed with isotypecontrol.

Example 7 Binding of Lag3 Clones to T Cells Isolated from Human andCynomolgous Monkey Blood

Binding of Lag3 mAb clones to primary T-cells was accessed using humanand cynomolgous monkey (cyno) peripheral blood mononuclear cells (PBMC).Human blood was obtained from the local donors; cyno blood was obtainedfrom Bioreclamation. PBMC were isolated from the whole blood usingFicoll-Paque Plus (GE Healthcare, Cat #17-1440-03) density gradientcentrifugation at 524×g for 40 minutes. PBMC were collected from themedium:plasma interface and washed 2 times with phosphate bufferedsaline (PBS). The residual red blood cells were lysed usingammonium-chloride-potassium red blood cell lysing solution (ACK, GIBCO,cat #A10492-010). Cynomolgus monkey or human PBMC (3×10⁶/ml) werestimulated with 4 μg/ml PHA (Sigma, L2769) for 48 h and 20 h,respectively. For flow cytometric analysis, 1×10⁶ PHA-activated PBMCswere used per staining in 50 μl FACS staining buffer (BD, cat #554657).Lag3 mAb clones or correspondent isotype controls (Table 11) wereincubated with human or cyno PBMS followed by detection using goatanti-mouse IgG-PE (BD 550589). Mouse anti-human CD3-pacific blue (BD558124, clone SP34-2), mouse anti-human CD4-PerCP (BD 550631, cloneL200), and mouse anti-human CD8-APC-Cy7 (BD 557834, clone SK1) were usedas phenotypic markers. Sample acquisition was performed on an LSR IIflow cytometer and the data were analyzed using FlowJo software version10.0.6 (Tree Star, Inc.).

Flow cytometry analysis revealed binding to all the analyzed Lag3 clonesto primary human and cyno T cells (representative data shown on FIG. 2).

TABLE 11 Lag 3 clones and isotype controls utilized in the studyTC31.3E1.C7 28ACM mIgG2a/K 10 mM NaPhosphate, 75 mM NaCl, 3% sucrose, pH7.3 TC31.27F11.C2 64AFW mIgG1/K 10 mM phosphate, 75 mM NaCl, 3% sucrosepH 7.4 KM9.LP1.MAB3 63ADP mIgG2b/K 20 mM NaAcetate, 7% sucrose, pH 5.5In house Lag3 lot# isotype formulation LB148.19E8.G1.1A1 42AGF mIgG2b/K10 mM PO4, 75 mM NaCl, 3% sucrose pH 7.4 LB145.22D2.E1.1D1 98AGFmIgG2a/K 10 mM PO4, 75 mM NaCl, 3% sucrose pH 7.4 LB148.4A10.1H1 45AGFmIgG2a/K 10 mM PO4, 75 mM NaCl, 3% sucrose pH 7.4 LB148.11C9.1C1 47AGFmIgG1/K 20 mM NaAc, 9% sucrose pH 5.0

Example 8 Epitope Mapping of hLAG3 Antibodies by Hydrogen DeuteriumExchange Mass Spectrometry

Contacts between anti-LAG3 antibodies and human LAG3 were determined byuse of hydrogen deuterium exchange mass spectrometry analysis.

Materials

-   -   Human LAG3-His:[VEGF leader]-LAG3_H-[His9G]

(SEQ ID NO: 447) (LQPGAEVPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRLRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDSGPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGRAPGALPAGHLLHHHHHHHHHGGQ)

-   -   Anti-hLAG3 clone 22D2 (chimeric antibody comprising the [LAG3_H]        mAb (LB145.22D2.E1.D1 (VHO/VLO) Geneart) IgG4 S228P/kappa (PX)        and the human IgG4 framework containing the hinge stabilizing        S228P mutation)    -   Anti-hLAG3 clone 11C9 (mouse×[LAG3_H] mAb (LB148.11C9.1C1)        IgG1/kappa (HY)/mouse IgG1)    -   Anti-hLAG3 clone 4A10 (mouse×[LAG3_H] mAb (LB148.4A10.1H1)        IgG2a/kappa (HY)/mouse IgG2a)        Liquid Chromatography-Mass Spectrometry

The mass spectrometer was a Thermo Scientific Orbitrap-Velos. For themeasurement of deuterium labeled samples, the mass spectrometer was setto acquire one full scan MS data in the orbitrap at 60,000 resolvingpower, a target ion count of 1E6 and a maximum ion injection time of 500millisecond. For the acquisition of MS/MS data for peptideidentifications, the mass spectrometer was set to acquire one full scanspectrum at 60,000 resolving power followed by ten data-dependent MS/MSspectra in the ion trap.

The liquid chromatography system was a Waters nanoAcquity for theanalytical column gradient and a Waters 515 isocratic pump for thesample digestion and loading. For sample digestion and loading, thebuffer used was 2% acetonitrile and 0.05% trifluoroacetic acid at a flowrate of 80 ul/min. For the analytical gradient, the buffers were BufferA) 0.1% formic acid in water and Buffer B) 0.1% formic acid inacetonitrile.

The gradient was at 40 ul/min from 2% B to 36% B in 10 minutes, followedby a wash of 80% B for 1.5 minute and a reequilibration at 2% B for 3minutes. The column was then washed by cycling the gradient between 2%and 80% B, three times with 1 minute at each step, followed by a finalequilibration at 2% B for 5 minutes. The trapping column was a WatersVanguard C18 BEH 1.7 um Guard Column and the analytical column was aWaters C18 BEH300, 1.7 um 1×50 mm column.

Sample handling for the deuterium labeling was done by a Leaptec H/D-XPAL system. The labeling sample tray was set to a temperature of 25° C.,the quenching tray was set to 1.5° C. and the trap and analytical columnchamber was set to 1.5° C. The immobilized pepsin column (PorosymeImmobilized Pepsin 2.1×30 mmm, Life Technologies) was kept outside thecolumn chamber at room temperature.

Deuterium Labeling

For the antibody clones 22D2 and 11C9, hLAG3-His (113 pmol/ul) was mixedwith an equal volume of the antibody (55 pmol/ul) or, as the unboundcontrol, PBS pH 7.6. The antibody bound samples and the unbound controlwere incubated at room temperature for 1 hour before beginning thelabeling experiment. For 4A10, the antibody (30 pmol/ul) was mixed withhLAG3-His at a 2:1 v/v ratio. Subsequent steps were identical for allthe samples.

To deuterium label the samples, 2 ul of sample was mixed with 25 ul ofPBS in deuterium oxide pD 7.6. Labeling time points were 30, 300, 3000or 9000 seconds. After the set time, 25 ul of the labeling mixture wasadded to 40 ul of cold quench buffer (8M Urea, 100 mM TCEP, 0.02%dodecylmaltoside). The quenched sample was incubated at 1.5° C. for 2minutes. 55 ul was then injected into the column cooling chamber wherethe sample was passed over the pepsin column and the resulting peptidesloaded onto the trapping column. After three minutes, the analyticalgradient and the mass spectrometer were started.

A fully deuterated sample was generated by incubating 2 ul of hLAG3 with108 ul of deuterated denaturing buffer (4M Urea, 100 mM TCEP, 0.01% DDMin 99.5% deuterium oxide). The sample was incubated at 37° C. overnight.55 ul was then directly injected into the column chamber and the dataacquired as before.

Data Analysis

LC-MS/MS data was acquired of an unlabeled sample and searched beforedeuterium labeling to verify successful digestion of the proteins and togenerate a list of peptides. Data was database searched using ProteomeDiscoverer 1.4 and the SEQUEST HT search algorithm (ThermoFisherScientific). The protein database used was the human LAG3-His sequenceconcatenated to the yeast Saccharomyces cerevisiae database.

MS data from the deuterium labeling experience was processed byHDExaminer (Sierra Analytics). The mass and retention time selected bythe software for each peptide was verified manually.

Results

The human LAG3 residues protected by the 22D2, 4A10 and 11C9 antibodies,when bound to the LAG3 protein, are illustrated in FIGS. 3, 4(a-c) and5(a-b).

Example 9 Cell Based KinExA

The affinities of the LAG3 antibodies for LAG3 were determined using acell based Kinetic Exclusion Assay (KinExA). Cell based KinExA can beused to measure the affinity of a molecule for a binding partner on acell surface (Rathanaswami et al. Analylitical Biochemistry 373(1):52-60 (2008); Xie et al. J. Immunol. Methods 304 (1-2): 1-14 (2005)). Inthis case, BaF/3 cells were stably transfected with human and cynomolgusmonkey LAG3 proteins. Transfected cells or the parental BaF/3 controlcell line were grown to a density of 1.7×10⁶ to 3.2×10⁶ cells per ml at37° C., 120 RPM, 5% CO₂, in 1× RPMI 1640 media with 10% FBS, 10 ng/mlIL-3, 5/ml puromycin. Cells were concentrated, mixed with 15 pM or 150pM antibody in cell culture media and incubated 24 to 48 hours at roomtemperature while rotating at 20 to 30 RPM. Cells were present at a topconcentration of 2×10⁷ cells per ml (parental BaF/3 or cynomolgus LAG3transfectants) or 1×10⁷ cells per ml (human LAG3 transfectants) anddiluted in a 2-fold, 18 member series. The cells were pelleted and freeantibody in the supernatant was measured using a KinExA 3200 instrument(Sapidyne, Idaho, USA). The instrument bound the free antibody topolymethyl methacrylate beads (Sapidyne) that had been coated with goatF(ab′)₂ anti-human Fcγ (Jackson ImmunoResearch Laboratories,Pennsylvania, USA). Antibody on the beads was labeled with 1.5 μg/mlAlexa Fluor® 647 conjugated goat anti-human (Fab′)₂ (JacksonImmunoResearch Laboratories), washed and the fluorescent signal was readall using the KinExA™ 3200. The data from the 15 pM and 150 pMconcentrations of each antibody were fit simultaneously using KinExA™Pro n-Curve Analysis software version 4.0.11 (Sapidyne).

TABLE 12 KinExA affinity measurements. human cynomolgus LAG3 K_(D) LAG3K_(D) K_(D) (pM) n K_(D) (pM) n 22D2 Chimera 3 2 11 2 Hu22D2 VH6/VL3 (Ab5) 6 2 25 2 Hu22D2 VH6/VL3 N55S (Ab 7) 10 2 11 2 Hu22D2 VH6/VL3 N55Q (Ab8) 11 2 16 2 Hu22D2 VH6/VL3 N55D (Ab 6) 2 3 12 3

Example 10 Effect of Anti-LAG3 Antibodies on Murine T-Cell Hybridoma 3A9Cells Expressing Human LAG3

The ability of anti-LAG3 antibodies to enhance antigen-specific T cellactivation was tested in a modified version of a previously described Tcell activation assay (Workman et al., Eur. J. Immunol. 32:2255-2263(2002)).

A HEL peptide₄₈₋₆₃-specific mouse T cell hybridoma (3A9) was stimulatedwith a haplotype-matched, MHCII⁺, HEL peptide₄₈₋₆₃-pulsed B cell line(LK35.2) and IL-2 release was assessed as a readout for antigen-specificT cell activation. The 3A9 T cell response to HEL peptide₄₈₋₆₃-pulsedLK35.2 cells was dose-dependent.

3A9 T cell lines stably overexpressing mouse or human LAG3 weregenerated by retroviral transduction. It was demonstrated that mouseMHC2 on LK35.2 cells can engage both human and mouse LAG3, resulting ina strong reduction of the antigen-specific IL-2 production, atsuboptimal T cell activation concentrations. The maximal effect of LAG3activity was observed when titrating HEL peptide₄₈₋₆₃ at a concentrationof 31.2 nM. The inhibitory effect of LAG3 overexpression was not seenwhen using LK35.2 B cells pulsed with higher peptide doses(corresponding to >100 nM). Treating with 10 ug/ml of a commerciallyavailable mouse LAG3 antibody, C9B7W, IL-2 levels were rescued to thatof the vector 3A9 cells.

To assess the effect of anti-LAG3 antibodies in this assay, mouse orhuman LAG3-overexpressing 3A9 T cells (100,000 per well) were pretreatedwith anti-LAG3 antibodies (serially diluted in 3-fold dilutions from 10ug/ml) for 30 minutes at 37° C., and stimulated with LK35.2 cells(33,333 per well) pulsed for 30 minutes prior to co-culture with 31.2 nMHEL peptide₄₈₋₆₃. After stimulation for 24 h at 37° C. and 5.0% CO₂,IL-2 secretion was assessed in culture supernatants by mesoscale.Inhibition of LAG3 with an antagonist antibody restored T-cell functionresulting in the rescue of IL-2 production in a dose-dependent manner.IL-2 production was not rescued when 3A9 cell were pre-treated withisotype control antibodies. The ability of LAG3 overexpression tosuppress IL-2 secretion coupled with IL-2 rescue after treatment withanti-LAG3 antibody validated this assay as a robust screening tool.Table 13 lists the EC₅₀s for IL-2 rescue using the hLAG3-3A9 system forthe mouse anti-human and humanized anti-LAG3 antibodies.

TABLE 13 Mouse anti-human LAG3 antibodies stimulate IL-2 production inthe hLAG3-3A9 T cell system Range EC50, nM Antibody (n)LB145.22D2.E1.1D1 1.06-1.65 (2) LB148.19E8.G1.1A1 1.74-1.83 (2)LB148.11C9.1C1 3.56-4.06 (2) LB148.4A10.1H1 2.83-2.96 (2) 22D2 chimera0.69-1.91 (5) Hu22D2 VH6/VL3 0.57-1.07 (Ab 5) (6) Hu22D2 VH6/VL30.45-1.27 N55S (Ab 7) (6) Hu22D2 VH6/VL3 0.47-1.01 N55D (Ab 6) (6)Hu22D2 VH6/VL3 0.72-1.08 N55Q (Ab 8) (6)

Example 11 Blocking of LAG3/MHC Class II Binding on Daudi Cells

Mouse anti-human LAG3 and humanized anti-LAG3 clones were tested fortheir ability to block hLAG3 interaction with human MHC Class II. Daudicells (ATCC #CCL-213) were used as a cell line positive for human MHCclass II expression. Daudi cells were blocked with 10 ug/ml of goat IgGin HBSS and 2% BCS on ice for 30 minutes and 0.5×10⁶ cells/sample werealiquoted into a 96-well V-bottom plate and blocking buffer removed.Clones LB145.22D2.E1.1D1, LB148.19E8.G1.1A1, LB148.4A10.1H1, andLB148.11C9.1C1 and hu22D2 VH6/VL3, hu22D2 VH6/VL3 N55Q, hu22D2 VH6/VL3N55S, hu22D2 VH6/VL3 N55D, and chimeric 22D2 were serially dilutedstarting at 20 ug/ml in HBSS/2% BCS and pre-incubated with 2 ug/ml ofhuman LAG3-huFc or biotinylated human LAG3-huFc in 96-well polypropyleneU-bottom plates in a final volume of 100 ul and incubated on ice for 30minutes. Following pre-incubation, the human LAG3-Fc/antibody antibodymixtures were added to the blocked Daudi cells and incubated for 45minutes on ice. Cells were pelleted by centrifugation at 1200 rpm andwashed twice with HBSS/2% BCS. Human LAG3-Fc binding to Daudi cells wasdetected using F(ab)′₂ goat anti-human IgG-PE conjugate (SouthernBiotech Cat #) at 1:200 for unconjugated huLAG3-huFc or 1:500 dilutionof streptavidin-PE for biotinylated huLAG3-huFc in 100 ul stainingvolume and incubated on ice for 20 minutes. Cells were washed twice asdescribed above, resuspended in HBSS/2% BCS and read on the FACSCalibur.Table 14 summarizes the IC50s for MEW class II blockade for the mouseanti-human LAG3 clones.

Humanized anti-human LAG3 antibodies (VH6/VL3, VH6/VL3 N55D, VH6/VL3N55Q, VH6/VL3 N55S, and chimeric 22D2) were tested for their ability toblock hLAG3 interaction with human MEW class II as described above.Biotinylated human LAG3-huFc was used and detection was usingstreptavidin-PE.

TABLE 14 Mouse anti-human LAG3 antibodies block the interaction of humanMHC Class II with human LAG3-Fc recombinant protein. Antibody IC50 (nM)LB145.22D2.E1.1D1 2.1 LB148.19E8.G1.1A1 2.8 LB148.11C9.1C1 2.0LB148.4A10.1H1 1.9 Chimeric 22D2 2.5 Hu22D2 VH6/VL3 2.6 (Ab 5) Hu22D2VH6/VL3 N55S 2.1 (Ab 7) Hu22D2 VH6/VL3 N55D 2.4 (Ab 6) Hu22D2 VH6/VL32.5 N55Q (Ab 8)

Example 12 Binding of Lag3 Clones to T Cells Isolated from Human andCynomolgous Monkey Blood

Binding of LAG-3 mAb clones to primary T-cells was assessed using humanand cynomolgous monkey (cyno) peripheral blood mononuclear cells (PBMC).Human blood was obtained from the local donors; cyno blood was obtainedfrom Bioreclamation. PBMC were isolated from the whole blood usingFicoll-Paque Plus (GE Healthcare, Cat #17-1440-03) density gradientcentrifugation at 524×g for 40 minutes. PBMC were collected from themedium:plasma interface and washed 2 times with phosphate bufferedsaline (PBS). The residual red blood cells were lysed usingammonium-chloride-potassium red blood cell lysing solution (ACK, GIBCO,cat #A10492-010). Cynomolgus monkey or human PBMC (3×10⁶/ml) werestimulated with 4 μg/ml PHA (Sigma, L2769) for 48 h and 20 h,respectively. For flow cytometric analysis, 1×10⁶ PHA-activated PBMCswere used per staining in 50 μl FACS staining buffer (BD, cat #554657).LAG-3 mAb clones or correspondent isotype controls were incubated withhuman or cyno PBMS followed by detection using goat anti-mouse IgG-PE(BD 550589). Mouse anti-human CD3-pacific blue (BD 558124, cloneSP34-2), mouse anti-human CD4-PerCP (BD 550631, clone L200), and mouseanti-human CD8-APC-Cy7 (BD 557834, clone SK1) were used as phenotypicmarkers. Sample acquisition was performed on an LSR II flow cytometerand the data were analyzed using FlowJo software version 10.0.6 (TreeStar, Inc.).

Flow cytometry analysis revealed binding of all the analyzed anti-LAG3antibodies to primary human and cyno T cells. EC₅₀ for binding to humanand cynomolgus CD4⁺ and CD8⁺ T cells were determined for humanized 22D2antibodies and are summarized in Table 15. The data represent 3 humanand 3 cynomolgus donors after stimulation with 4 ug/ml of PHA for 40hours prior to staining.

TABLE 15 EC₅₀ for binding to human and cynomolgus CD4⁺ and CD8⁺ T cells.(EC50, pM) Human Cynomolgus LAG-3 LAG-3 CD4⁺ CD8⁺ CD4⁺ CD8⁺ Hu22D2VH6/VL3 N55S 57 49 35 41 (Ab 7) Hu22D2 VH6/VL3 N55D 39 33 30 30 (Ab 6)Hu22D2 VH6/VL3 N55Q 41 35 27 31 (Ab 8)

Example 13 Effect of Anti-Human LAG-3 Antibody Treatment+1-Anti-PD-1Treatment on IL-2 Production in SEB Stimulated Human PBMCs

Human primary T cell assays—The first assay tested the function ofblocking LAG3 alone or in combination with anti-PD-1 to increase IL-2production by T cells following SEB activation of human PBMCs (FIG. 7).Neutralization of LAG3 activity, alone and in the presence of anti-PD-1,resulted in enhanced IL-2 production. Examples of the IL-2 mesoscalecounts from 2 donors across a dose titration of anti-LAG3 antibody areshown. Hu22D2-enhanced IL-2 production by SEB-stimulated PBMC by1.54-fold (range 1.15-2.36-fold, n=8) compared to isotype control and incombination with anti-PD1 by 1.45 fold (range 1.15-2.36-fold, n=4 at 10or 0.3 μg/ml of anti-PD-1) as compared to anti-PD-1 alone plus isotypecontrol. Hu22D2 alone or in combination with anti-PD-1, showedcomparable activity to a benchmark anti-LAG3 antibody. Hu2D2 is Ab6.α-PD-1 is a fully human IgG4 anti-human PD1 monoclonal antibody.Benchmark anti-LAG3 is a fully human IgG4 anti-human LAG3 monoclonalantibody that binds to the human LAG3 extraloop.

Example 14 Effect of hu22D2 Treatment+1-Anti-PD-1 Treatment on IFN-Gammaand TNFα Production in MLR Stimulated Human PBMCs

A MLR system was also used to test the activity of hu22D2 in primary Tcells. Human PBMCs are stimulated with monocyte-derived dendritic cellsfrom a different donor, leading to T cell activation due to MEWmismatch. A high degree of variability in donor responses was observedin this assay. However, preliminary data demonstrated thatneutralization of LAG3 activity, alone and in the presence of anti-PD-1enhanced T cell activation in two out of three donors, as assessed byIFN-γ and TNF-α production (FIG. 8). Hu2D2 is Ab6.

Example 15 Pharmacokinetic and Pharmacodynamics Analysis of Anti-LAG3 inCynomolgous Monkeys

Pharmacokinetic and pharmacodynamics profile of Ab6 in cynomolgousmonkeys was evaluated. Procedures involving the care and use of animalsin the study were reviewed and approved. During the study, the care anduse of animals were conducted in accordance with the principles outlinedin the guidance of the Association for Assessment and Accreditation ofLaboratory Animal Care (AAALAC), the Animal Welfare Act, the AmericanVeterinary Medical Association (AVMA) Euthanasia Panel on Euthanasia,and the Institute for Laboratory Animal Research (ILAR) Guide to theCare and Use of Laboratory Animals.

25 naïve male cynomolgus monkeys of Chinese origin (4.0-7.0 kg at timeof dosing) were used. Animals were observed twice daily. Additionally,animals were observed at each blood collection time point. Body weightswere recorded once prior to each dosing occasion.

In the study, five males were assigned into each of 5 groups. Animals inall groups were administered 5 doses of test or control articles IV viaa cephalic vein over 10 minutes. The doses for the groups were 0.03mg/kg, 0.3 mg/kg, 1 mg/kg, 10 mg/kg and 30 mg/kg. Pharmacokineticsamples were drawn for all animals on Day 1: predose and 15 minutes and1, 4, 8, 24 (Day 2), 48 (Day 3) and 96 (Day 5) hours post dose. Sampleswere also collected on Day 8: predose and 1, 8, 24 (Day 9), 48, (Day10), 120 (Day 13) and 168 (Day 15) hours post dose. All samples wereprocessed to plasma, stored frozen at −70° C.±10° C. until analyzed.Pharmacodynamic samples were drawn for all animals on Day 1: predose and15 minutes and 1, 4, 8, 24 (Day 2), 48 (Day 3) and 96 (Day 5) hours postdose. Samples were also collected on Day 8: predose and 1, 8, 24 (Day9), 48 (Day 10), 120 (Day 13) and 168 (Day 15) hours post dose. Allsamples were processed to plasma, stored frozen at −70° C.±10° C. untilanalyzed.

Total antibody and free (unbound antibody and partially bound) wereevaluated by two different assays, universal assay and antigen captureassay, respectively. The data from this evaluations were set forth inFIG. 9 (Ag-Capture assay results) and FIG. 10 (Universal Assay results).The clearance and volume of distribution at steady state for each doselevel were estimated and tabulated in Tables 16 (antigen capture assay)and 17 (universal assay). The differences between the antibody clearanceand volume of distribution parameters demonstrated that a portion of theAb6 antibody was engaging its target in the cynomolgous monkey subjects.

TABLE 16 LAG3 antigen-capture assay-Clearance of Ab6 and volume ofdistribution at steady state (Vss) at various doses. Dose Clearance Vss(mg/kg) (mL/hr/kg) (mL/kg) 0.03 6.8 76.4 0.3 0.48 75.5 1 0.45 95.3 100.33 72.3 30 0.29 63.6

The antigen-capture data detected free antibody in the sample. Theseantigen-capture data (data not shown) demonstrated a 23-fold differencein clearance over the dose range investigated (0.003 mg/kg-30 mg/kg)(Table 16).

TABLE 17 Total antibody assay-Clearance of Ab6 and volume ofdistribution at steady state (Vss) at various doses. Dose Clearance Vss(mg/kg) (mL/hr/kg) (mL/kg) 0.03 0.94 47 0.3 0.36 48.9 1 0.33 56.1 100.26 60.9 30 0.26 66.5

The total antibody data detected total antibody in the sample. Thesetotal antibody data (FIG. 9) demonstrated a 3.6-fold difference inclearance over the dose range investigated (0.003 mg/kg-30 mg/kg) (Table17).

The total concentration of Ab6 in cynolmolgous monkeys at the 0.03mg/kg, 0.3 mg/kg, 1 mg/kg, 10 mg/kg and 30 mg/kg doses over time wereevaluated. The data from this evaluation are set forth in FIG. 10. Asthe dose increased, the clearance of the antibody from the subjectsdecreased. The dose-normalized concentration data of Ab6 over time isset forth in FIG. 11. At later timepoints, after the second IV bolusadministration of Ab6, there was loss of exposure indicating potentialimmunogenicity against the antibody.

LAG3 target-mediated clearance of the Ab6 antibody was observed whenconcentration of the antibody were observed in the monkey subjects overtime. A two compartment PK model with linear and non-linear (Vmax, Km)clearance parameters was developed to describe the concentration-timeprofiles for Ab6. These data are set forth in FIG. 12.

All references cited herein are incorporated by reference to the sameextent as if each individual publication, database entry (e.g. Genbanksequences or GeneID entries), patent application, or patent, wasspecifically and individually indicated to be incorporated by reference.This statement of incorporation by reference is intended by Applicants,pursuant to 37 C.F.R. § 1.57(b)(1), to relate to each and everyindividual publication, database entry (e.g. Genbank sequences or GeneIDentries), patent application, or patent, each of which is clearlyidentified in compliance with 37 C.F.R. § 1.57(b)(2), even if suchcitation is not immediately adjacent to a dedicated statement ofincorporation by reference. The inclusion of dedicated statements ofincorporation by reference, if any, within the specification does not inany way weaken this general statement of incorporation by reference.Citation of the references herein is not intended as an admission thatthe reference is pertinent prior art, nor does it constitute anyadmission as to the contents or date of these publications or documents.

The present invention is not to be limited in scope by the specificembodiments described herein. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and theaccompanying figures. Such modifications are intended to fall within thescope of the appended claims.

The foregoing written specification is considered to be sufficient toenable one skilled in the art to practice the invention. Variousmodifications of the invention in addition to those shown and describedherein will become apparent to those skilled in the art from theforegoing description and fall within the scope of the appended claims.

We claim:
 1. A polynucleotide encoding a light chain variable domain, aheavy chain variable domain, or both a light chain variable domain and aheavy chain variable domain of an antibody or antigen binding fragmentthat specifically binds to human Lymphocyte Activation Gene-3 (LAG3),wherein the light chain variable domain comprises: CDR-L1 that comprisesthe amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); andthe heavy chain variable domain comprises: CDR-H1 that comprises theamino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprises theamino acid sequence: DINPNX₁GGTIYX₂QKFX₃X₄ (SEQ ID NO: 446), wherein,X₁=D, N, S or Q X₂=A or S X₃=Q or K, and X₄=E or G; and CDR-H3 thatcomprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35).
 2. Thepolynucleotide of claim 1, wherein the antibody or antigen-bindingfragment comprises: (1) a light chain variable domain comprising: CDR-L1that comprises the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38);CDR-L2 that comprises the amino acid sequence: GASNLES (SEQ ID NO: 39);and CDR-L3 that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO:40); and a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNNGGTIYAQKFQE (SEQ ID NO: 458); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or(2) a light chain variable domain comprising: CDR-L1 that comprises theamino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 thatcomprises the amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and aheavy chain variable domain comprising: CDR-H1 that comprises the aminoacid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprises the aminoacid sequence: DINPNSGGTIYAQKFQE (SEQ ID NO: 456); and CDR-H3 thatcomprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (3) alight chain variable domain comprising: CDR-L1 that comprises the aminoacid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprisesthe amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 thatcomprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and aheavy chain variable domain comprising: CDR-H1 that comprises the aminoacid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprises the aminoacid sequence: DINPNDGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3 thatcomprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (4) alight chain variable domain comprising: CDR-L1 that comprises the aminoacid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprisesthe amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 thatcomprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and aheavy chain variable domain comprising: CDR-H1 that comprises the aminoacid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprises the aminoacid sequence: DINPNQGGTIYAQKFQE (SEQ ID NO: 455); and CDR-H3 thatcomprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35); or (5) alight chain variable domain comprising: CDR-L1 that comprises the aminoacid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38); CDR-L2 that comprisesthe amino acid sequence: GASNLES (SEQ ID NO: 39); and CDR-L3 thatcomprises the amino acid sequence: QQSTEDPRT (SEQ ID NO: 40); and aheavy chain variable domain comprising: CDR-H1 that comprises the aminoacid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprises the aminoacid sequence: DINPNGGGTIYAQKFQE (SEQ ID NO: 454); and CDR-H3 thatcomprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35).
 3. Thepolynucleotide of claim 1, wherein the antibody or antigen-bindingfragment comprises: a light chain variable domain comprising: CDR-L1that comprises the amino acid sequence: KASQSLDYEGDSDMN (SEQ ID NO: 38);CDR-L2 that comprises the amino acid sequence: GASNLES (SEQ ID NO: 39);and CDR-L3 that comprises the amino acid sequence: QQSTEDPRT (SEQ ID NO:40); and a heavy chain variable domain comprising: CDR-H1 that comprisesthe amino acid sequence: DYNVD (SEQ ID NO: 33); CDR-H2 that comprisesthe amino acid sequence: DINPNQGGTIYAQKFQE (SEQ ID NO: 457); and CDR-H3that comprises the amino acid sequence: NYRWFGAMDH (SEQ ID NO: 35). 4.The polynucleotide of claim 1, wherein the antibody or antigen-bindingfragment comprises: (1) a light chain variable domain comprising theamino acid sequence set forth in amino acids 21-131 of SEQ ID NO: 126,and a heavy chain variable domain comprising the amino acid sequence setforth in amino acids 1-119 of SEQ ID NO: 106; or (2) a light chainvariable domain comprising the amino acid sequence set forth in aminoacids 21-131 of SEQ ID NO: 126, and a heavy chain variable domaincomprising the amino acid sequence set forth in amino acids 1-119 of SEQID NO: 108; or (3) a light chain variable domain comprising the aminoacid sequence set forth in amino acids 21-131 of SEQ ID NO: 126, and aheavy chain variable domain comprising the amino acid sequence set forthin amino acids 1-119 of SEQ ID NO: 110; or (4) a light chain variabledomain comprising the amino acid sequence set forth in amino acids21-131 of SEQ ID NO: 126, and a heavy chain variable domain comprisingthe amino acid sequence set forth in amino acids 1-119 of SEQ ID NO:112; or (5) a light chain variable domain comprising the amino acidsequence set forth in amino acids 21-131 of SEQ ID NO: 126, and a heavychain variable domain comprising the amino acid sequence set forth inamino acids 1-119 of SEQ ID NO:
 122. 5. The polynucleotide of claim 1encoding a light chain, a heavy chain, or both a light chain and a heavychain of an antibody that comprises: (1) a light chain immunoglobulincomprising the amino acid sequence set forth in amino acids 21-238 ofSEQ ID NO: 126, and a heavy chain immunoglobulin comprising the aminoacid sequence set forth in SEQ ID NO: 106; or (2) a light chainimmunoglobulin comprising the amino acid sequence set forth in aminoacids 21-238 of SEQ ID NO: 126, and a heavy chain immunoglobulincomprising the amino acid sequence set forth in SEQ ID NO: 108; or (3) alight chain immunoglobulin comprising the amino acid sequence set forthin amino acids 21-238 of SEQ ID NO: 126, and a heavy chainimmunoglobulin comprising the amino acid sequence set forth in SEQ IDNO: 110; or (4) a light chain immunoglobulin comprising the amino acidsequence set forth in amino acids 21-238 of SEQ ID NO: 126, and a heavychain immunoglobulin comprising the amino acid sequence set forth in SEQID NO: 112; or (5) a light chain immunoglobulin comprising the aminoacid sequence set forth in amino acids 21-238 of SEQ ID NO: 126, and aheavy chain immunoglobulin comprising the amino acid sequence set forthin SEQ ID NO: 114; or (6) a light chain immunoglobulin comprising theamino acid sequence set forth in amino acids 21-238 of SEQ ID NO: 126,and a heavy chain immunoglobulin comprising the amino acid sequence setforth in SEQ ID NO: 116; or (7) a light chain immunoglobulin comprisingthe amino acid sequence set forth in amino acids 21-238 of SEQ ID NO:126, and a heavy chain immunoglobulin comprising the amino acid sequenceset forth in SEQ ID NO: 118; or (8) a light chain immunoglobulincomprising the amino acid sequence set forth in amino acids 21-238 ofSEQ ID NO: 126, and a heavy chain immunoglobulin comprising the aminoacid sequence set forth in SEQ ID NO: 120; or (9) a light chainimmunoglobulin comprising the amino acid sequence set forth in aminoacids 21-238 of SEQ ID NO: 126, and a heavy chain immunoglobulincomprising the amino acid sequence set forth in SEQ ID NO:
 122. 6. Thepolynucleotide of claim 1, wherein the antibody or antigen-bindingfragment is humanized.
 7. The polynucleotide of claim 2, wherein theantibody or antigen-binding fragment is humanized.
 8. The polynucleotideof claim 3, wherein the antibody or antigen-binding fragment ishumanized.
 9. A polynucleotide encoding a light chain variable domain, aheavy chain variable domain, or both a light chain variable domain and aheavy chain variable domain of an antibody or antigen binding fragmentthat specifically binds to human Lymphocyte Activation Gene-3 (LAG3),wherein the light chain variable domain comprises amino acids 21-131 ofSEQ ID NO: 126 and the heavy chain variable domain comprises amino acids1-119 of SEQ ID NO:
 116. 10. The polynucleotide of claim 9 encoding alight chain, a heavy chain, or both a light chain and a heavy chain ofthe antibody, wherein the light chain comprises the amino acid sequenceset forth in amino acids 21-238 of SEQ ID NO: 126 and the heavy chaincomprises the amino acid sequence set forth in amino acids 1-119 of SEQID NO:
 116. 11. A polynucleotide encoding a light chain, a heavy chain,or both a light chain and a heavy chain of an antibody that specificallybinds to human Lymphocyte Activation Gene-3 (LAG3), wherein the lightchain comprises the amino acid sequence set forth in amino acids 21-238of SEQ ID NO: 126 and the heavy chain comprises the amino acid sequenceset forth in SEQ ID NO:
 116. 12. An expression vector comprising thepolynucleotide of claim
 1. 13. An expression vector comprising thepolynucleotide of claim
 2. 14. An expression vector comprising thepolynucleotide of claim
 9. 15. An expression vector comprising thepolynucleotide of claim
 10. 16. An expression vector comprising thepolynucleotide of claim
 11. 17. A host cell comprising the expressionvector of claim
 12. 18. A host cell comprising the expression vector ofclaim
 13. 19. A host cell comprising the expression vector of claim 14.20. A host cell comprising the expression vector of claim
 15. 21. A hostcell comprising the expression vector of claim 16.